Preparation and transformation of Saccharomyces cerevisiae receptor cells
Saccharomyces cerevisiae, also known as baker's yeast or budding yeast. Saccharomyces cerevisiae is the yeast most widely associated with humans, not only becau
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用于检测、定量和定位抗原与抗体的技术——ELISA、免疫印迹、免疫组化与流式细胞术。以下是我们知识库中标注该主题的实验方案、常见问题与技术文章。
219 项资源 · 实验方案、常见问题与文章
Preparation and transformation of Bichler's yeast receptor cells
The most commonly used transformation methods for Saccharomyces cerevisiae are the electrotransformation method and the protoplast method, among which the elect
Hybridization and gene localization experiments in Escherichia coli
The E. coli chromosome is ring-shaped. High-frequency recombinant strains (Hfr) have the F factor integrated into their chromosomes, and the chromosomes of the
Embryo cryopreservation
When embryo freezing is used to cryopreserve living tissues and cells, the freezing process will likely cause damage to the cells.
Embryo transfer in cattle
Embryo transfer is a necessary technical means and link in the research and development of embryo biotechnology, such as cloning, transgenic, in vitro fertiliza
in vitro transcription technology
Usually transcription occurs in the organism, if we will contain RNA transcriptase, NTP and other conditions, in an in vitro cell-free system, using DNA as a te
Indirect immunofluorescence of living cells
The surface of living cells retains a more complete antigen or receptor, first use specific mouse-derived monoclonal antibody to bind to the corresponding antig
Chip Preparation Techniques for Direct Dot Sampling
The direct dispensing method, first developed by Brown Laboratory of Stanford University, involves arranging trace oligonucleotide fragments, cDNA or proteins,
Isolation and exchange of Streptomyces crassus
Neurospora crassa (2n = 14), also known as red bread mold, taxonomically belongs to the Ascomycetes, coccolithophores, and pulsatilla genera of fungi, and four
Identification of Coumarin Glycosides
Coumarona odoruta was first found in the legume family Coumaraceae, so it is called coumarin, with a special aroma. Coumarins and sugars combined into a glycosi
Experimental modeling of heterologous in vitro fertilization
Heterozygous in vitro fertilization experimental modeling can be used to: (1) simulate in vivo conditions of male and female gametes from different species of a
Purification of antibodies from immune sera
Purified antibodies are often used for direct detection of antigens. In this case the antibody is labeled with an easily identifiable marker for binding and det
Dry Heat Sterilization Experiment
Dry heat sterilization is the use of high temperatures to sterilize microorganisms by coagulating and denaturing the proteins in their cells. It is used to (1)
Measurement of ammonia nitrogen in water
Ammonia nitrogen (NH3-N) is present in water as either free ammonia (NH3) or ammonium salts (NH4"), and the ratio of the two depends on the pH of the water and
Experiments for the determination of chemical oxygen demand in water samples
Chemical oxygen demand (Chemical oxygen demand, referred to as COD), refers to the amount of oxidant consumed when treating water samples with potassium dichrom
Measurement of suspended solids in wastewater
Suspended solid (Sespended solid, abbreviated as SS) refers to the solid material suspended in water, including inorganic and organic matter insoluble in water,
hemagglutination inhibition test
Viruses with hemagglutinin (HA) can agglutinate human or animal erythrocytes, which is known as hemagglutination phenomenon. Hemagglutination phenomenon can be
Novel vector-mediated gene introduction technologies
Currently gene introduction technology is the main technique for the preparation of ips cells, and the improvement of gene vectors promotes the establishment of
Experiments on the preparation of suppositories
Suppositories can be categorized into two types according to their action. One is localized in the lumen; the other is absorbed from the lumen into the bloodstr
Water pH Measurement Experiment
The pH value (pH value) of natural water is mostly in the range of 6 to 9, which is also the pH control range in China's wastewater discharge standards. pH valu
Red Wine Vinification
Yeast is categorized into two types: natural yeast and artificially cultured yeast. Natural yeasts are wild yeasts that are often attached to the fruit skin. Fr
Measurement of dissolved oxygen in water
This experiment is mainly used to determine dissolved oxygen in water.
Bacillus subtilis fermentation culture experiment
Starting with test tube strains, the culture is added by expanding step by step, eventually fermenting Bacillus subtilis using a mechanical mixing tank. Shaking
Experiment on the determination of water chromaticity
Clean water should be colorless at shallow depths and light blue-green at deeper depths. The presence of humus, mud, plankton, metal ions such as iron and manga
Measurement of nitrate nitrogen in water
Nitrate in water (Nitrate) is under aerobic conditions, nitrite nitrogen, ammonia nitrogen and other forms of nitrogen compounds in the most stable nitrogen com
Water conductivity measurement experiment
Electrical conductivity (Electric conductivity) is the ability of a solution to conduct an electric current expressed as a number. Pure water has a small conduc
Hybridoma cloning
Hybridoma cloning is mainly used to (1) clone antibody-positive holes. (2) Separating antibody-secreting cells, antibody-nonsecreting cells, desired antibody (s
Experiments on the preparation of membrane agents
Membrane agent refers to the membrane preparation made of drug and suitable film-forming material through processing for oral or mucosal external use. The resea
Wastewater Turbidity Measurement Experiment
Turbidity (Turbidity) is caused by the presence of sediment, clay, organic matter, inorganic matter, plankton and microorganisms and other suspended substances
Crystallization of lipid mesoporous membrane proteins
Crystallization of proteins using lipid mesoporous membrane proteins can be used to resolve X-ray crystal structures of prokaryotic and eukaryotic proteins, pro
Acetaminophen tablets solubility determination experiment
Dissolution degree refers to the speed and degree of dissolution of drugs from solid preparations such as tablets or capsules in the specified solvent. However,
Preparation of red currant pigment of currant rice
Red pigment is a red natural pigment produced by Aspergillus erythropolis during its growth and metabolism. Aspergillus erythropolis is currently the only micro
Experiments on the preparation of colloidal solution-type liquid pharmaceuticals
Colloidal solution type liquid pharmaceuticals according to the classification of the dispersion system, including two categories: a class of polymer solutions
Atmospheric Total Suspended Particulate Matter Measurement Laboratory
Total suspended particulate (TSP) is particulate matter suspended in the air with an aerodynamic equivalent diameter of ≤100 μm. It originates from solid or l
Measurement of suspended solids in water
For the food industry, by the environmental impact of the biggest factor is water, while food processing and pollution of the water environment, to prevent and
Guidance and recording of potentials of sciatic nerve trunk action
The experimental method was obtained from the official website of the Fourth Military Medical University
Measurement of chloride ions in water
Chloride ion measurement tests in water can be used to: test the quality of water, which, when high, can damage metal pipes and structures and prevent plant gro
Experiment for determination of total solids in water
Total solids in water (Total solid, referred to as TS), also known as the total evaporation residue, is in the prescribed conditions, water samples evaporation
Measurement of formaldehyde and ammonia in indoor air
Formaldehyde is a colorless, irritating and water-soluble gas that binds to proteins. Inhalation of high concentrations of formaldehyde can cause severe irritat
Measurement of fluoride ion in water
Fluoride (F-) is one of the necessary trace elements for human body, and lack of fluoride is prone to dental caries. Fluoride content in water is one of the imp
Cadmium, Copper, Zinc, Lead in Water Measurement Test
A water sample or an ablated water sample is drawn directly into a flame, and the atomic vapors formed in the flame absorb the characteristic electromagnetic ra
Experimental determination of chromium in soil by colorimetric method of dibenzoyl dihydrazide
For food raw materials, soil pollution is an important factor affecting the yield, quality and even food safety of raw materials, especially the heavy metal con
Activated carbon adsorption experiment
When the activated carbon adsorbs impurities in water, adsorption and desorption phenomena will occur at the same time, when the adsorption and desorption are i
Experiments on neutralization titration and determination of the ionization constant of acetic acid
This experiment is from the official website of Southern Medical University
Cadmium, Copper, Zinc and Lead Determination Experiment in Soil
The soil sample is digested and the solution is introduced into the atomizer, the metal atoms formed absorb the characteristic electromagnetic radiation, and th
Experiment for determination of total hardness of water
Total hardness (Total hardness) is the total concentration of calcium and magnesium ions in water and converted to calcium oxide for calculation. Hardness has a
Odor and taste checking experiments
For the food industry, by the environmental impact of the biggest factor is water, while food processing and pollution of the water environment, in order to pre
Experiments for the determination of water coloration
For the food industry, by the environmental impact of the biggest factor is water, while food processing and pollution of the water environment, to prevent and
coagulation experiment
Colloidal particles in the water are small or charged and stable in the water and difficult to settle, can be added to the coagulant in the water, destroying th
Effect of load on muscle contraction
The experimental method was obtained from the official website of the Fourth Military Medical University
Determination of the conduction velocity and duration of the action potential of the sciatic nerve trunk
The experimental method was obtained from the official website of the Fourth Military Medical University
Atmospheric sulfur dioxide measurement experiment
Sulphur dioxide is a typical air pollutant among sulphur-containing compounds, which originates from the combustion of fuels such as coal and petroleum, smeltin
Observation experiments on the basic morphological structures of pathogenic organisms
Microbial cells are small and transparent, not easy to identify under the ordinary optical microscope, they must be stained, so that the stained organisms and t
Experiments for the determination of total dissolved solids
After the water sample is filtered and dried at a certain temperature, the solid residue obtained is called total dissolved solids, including non-volatile solub
Measurement of calcium and magnesium in water
The specimens were drawn into an air-acetylene flame to atomize calcium and magnesium, and the absorption of the 422.7 nm resonance line was chosen to quantify
Biochemical Oxygen Demand Measurement Laboratory
Biochemical oxygen demand (Biochemical oxygen demand, referred to as BOD) refers to the conditions in the presence of oxygen, microbial decomposition of organic
Permanganate Index Measurement Experiment
Permanganate value (Permanganate value) is the amount of potassium permanganate consumed in treating water samples in acidic or alkaline media with potassium pe
Measurement of hexavalent chromium in water
The common valence states of chromium compounds are trivalent and hexavalent. In water bodies, hexavalent chromium generally exists in the form of CrO42-, Cr2O7
Atmospheric NOx Measurement Experiment
Nitric oxides (Nitric oxides) in the air are NO, NO2, N2O, N2O3, N2O4, N2O5, etc., of which NO and NO2 are the main forms. Nitrogen oxides are generally generat
Single and tonic contractions of muscles
Single and tonic contractions of a muscle can be used to observe the phenomenon of contractile response of a muscle to a stimulus.
Morphological observation experiment of Toxoplasma gondii
This experiment is from the official website of Parasitology, School of Medicine, Shandong University.
Experiments on the determination of fluoride content in agricultural products by fluoride ion selective electrode method
Fluorine is an essential nutrient for the human body and a toxic pollutant. Fluorine in nature mainly exists in natural minerals such as fluorspar and apatite.
High-throughput holistic in situ hybridization for the detection of environmental disturbances in zebrafish (Danio rerio), an embryonic tissue-specific gene expression alteration experiment
Holistic in situ hybridization is a process that allows observation of cellular gene expression (mRNA) in intact organisms. By comparing differences in gene exp
Application of suppression subtractive hybridization to detect metagenomic diversity in environmental samples
Metagenomics provides information on the total genetic structure and functional composition of nondisposing samples of microbial environments without the need t
Experimental purification of λ phage particles by precipitation/centrifugation
If the purpose of purification of the phage pellet is to obtain outgrown DNA fragments for subsequent subcloning or to prepare phage two-arms, centrifugation ca
Isolation, characterization and transplantation experiments of neural stem cells
Neural cells in the mammalian central nervous system (CNS) are derived from proliferating pluripotent stem cells or precursor cells that migrate, localize, and
Experiments on the resolution of electromyographic signals in muscle fibers
The precise EMG analysis techniques covered in this chapter recover all available information in the EMG signal.EMG signal analysis consists of two categories:
magnetoencephalography
In physics, any electric current is accompanied by the production of a magnetic field. Ion currents flowing inside the brain and nerves also generate magnetic f
Experimental molecular haplotype analysis method based on emulsion polymerase chain reaction
Genotypes can be assayed by a variety of experimental methods, but assays for haplotype analysis, such as molecular haplotype analysis, are limited. Dependent o
Extracellular matrix (ECM) preparation experiments
Experiments in the preparation of extracellular matrix can be used to (1) study the constituents of the extracellular matrix as well as its physiological charac
Handling of Petri dishes and plates
The manipulation of Petri plates is necessary for cell experiments such as (1) cell growth (2) cell propagation (3) cell inoculation.
Laboratory methods for the detection of chromosomal structural aberrations in human and mouse spermatozoa by fluorescence in situ hybridization
This new generation of sperm F I S H assays has identified multiple paternal risk factors such as age, polypharmacy, lifestyle, and environmental aspects and oc
Human nerve fiber microelectrode recording experiment
The purpose of this chapter is first to introduce the technique to scientists who wish to use it in a practical way. Secondly, typical examples of the results o
Electrical activity of single neurons: the membrane clamp technique
Over the last 20 years, the membrane clamp technique has become one of the main research tools in modern electrophysiology. Initially used only to measure singl
Chemical and protein hydrolysis modification experiments of antibodies
This chapter is intended as a practical guide for researchers who need to modify antibodies themselves. It covers common chemical and protein hydrolysis modific
Improvement of the Koch method of acute toxicity testing
1, study the design of acute toxicity test of chemicals, master the determination of LD50. 2. Observe the toxic reaction of phoxim.
Determination of nitrate in agricultural products
The experiment for the determination of nitrate in agricultural products can be used for: (1) detecting the content of nitrate and nitrite in food; (2) as one o
Restriction enzyme digestion labeling genome scanning for mutation detection experiments
Today, there is a need for more effective methods in the detection of mutations in humans and other organisms caused by environmental toxins and radiation expos
Fluoride Detection in Agricultural Products by Ashing Experiment
Fluorine is an essential nutrient for the human body and a toxic pollutant. Fluorine in nature mainly exists in natural minerals such as fluorspar and apatite.
Optical recording experiments of population neurons in brain slices
Optical recording involves the use of molecular indicators whose optical properties (absorption or fluorescence) vary according to the parameters of cellular ac
Central nervous system culture techniques: hands-on experiments in chick embryo neuron culture
Establishing an accurate and reusable neuronal culture model in vitro is of self-evident importance for experimental research in neuroscience. Many research gro
Experiments on collection and preparation of biological materials for laboratory animals
The study of the toxic effects of foreign compounds. It is often necessary to determine the concentrations of compounds or their metabolites in the blood, urine
Analytical methods for correlating electrical muscle activity with animal locomotion
This chapter deals with two main aspects: (i) the methodology for simultaneous recording of EMG and video on normally moving animals, and (ii) the timing and ne
Ultra-clean table aseptic operation method
Clean and wipe the work area, and then place bottles, pipettes, etc. in the work area. Then, start the preparatory work of preparing the culture medium and othe
Operating on an open workbench
Clean and wipe the work area, place the reagent bottles, pipette, etc., on the bench (Figure 6.5), and make preparations. If necessary. If necessary, flame ste
Laboratory animal grouping, labeling, and dyeing experiments
The grouping, labeling and reasonable dyeing of experimental animals are the prerequisites for obtaining good experimental results and conclusions, and are also
Determination of nitrite in agricultural products
The level of nitrate and nitrite in food is one of the most important pollutants that affect food quality and jeopardize human health. Its content in food mater
Experiments on optical recording techniques for culturing single neurons
Techniques for optically recording the course of activity of individual viable neurons must be considered in the light of two fundamental questions, namely, wha
positron emission tomography (PET)
Positron emission tomography (PET) is an in vivo functional neuroimaging technique that provides three-dimensional quantitative images of a wide range of physio
Fluorescein in situ hybridization for detection of chromosomal aberrations and aneuploidies caused by environmental toxins
Many chemical agents and ionic radiation of different nature can cause damage to chromosome integrity, resulting in abnormal chromosome distribution in mitosis
Site-specific integration experiments mediated by bacteriophage ΦC31 integrase
Efficient delivery of ΦC31 integrase and donor plasmids to recipient tissues or cells of interest remains the most challenging element of this integrative syst
Gene gun technology for delivering genes into the skin
Gene gun technology is a fast and simple technique for delivering genes into cells. This technology has many advantages: it allows direct delivery of plasmids w
Polylysine copolymers for gene delivery
Polylysine and its copolymers have gained wide application as non-viral gene vectors. Although polylysine itself is toxic to cells, the covalent attachment of p
Recombinant herpes simplex virus vector
The natural physiological characteristics of H S V -I include the establishment of a lifelong latent state in neurons, where the viral gene remains a free molec
technology that targets genes
Gene pooping is an experimental tool for targeted alteration of genetic information of a species based on embryonic stem cells (ESCs) and homologous recombinati
Design method for adeno-associated heterotrimeric viral vectors
A A V coat peptide display library, error-prone P C R and D N A recombination . A A V serotypes with different histophilicity can be generated by natural mutati
Experiments on Prokaryotic Microinjection Technique
A technique for preparing transgenic mice by prokaryotic microinjection is described. Prokaryotic microinjection, which involves the injection of DNA into haplo
Human brain invasive techniques: microelectrode recording and microstimulation
The techniques discussed in this chapter allow recording under local anesthesia of the activity of cortical and subcortical structures such as the thalamus, bas
Labeling experiments with fluorescence in situ hybridization probes for genomic target sequences
Fluorescence in situ hybridization (FISH) requires nucleic acid probes, including deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or nucleic acid analogs t
Preservation and amplification of targeted modified measles viruses
The S T A R System (6 Histidine Labeling and Retargeting) employs a pseudo-receptor and a label for a virus to enable the rescue and amplification of fully reco
Reverse transcription virus vector assay
Reverse transcription viruses are well suited to be used as gene introduction tools because of their specific life cycle and ability to integrate efficiently in
Non-viral gene transfer experiments with Trojan horse liposomes that can cross the blood-brain barrier
Non-viral vector plasmid DNA can be delivered to the human brain via the vascular route across the blood-brain barrier by intravenous injection of DNA encapsula
Cyclodextrin-containing polycations for nucleic acid delivery
A large number of non-viral systems have been used for nucleic acid delivery to cultured cells in vitro or to specific cells in vivo. They vary considerably in
Experiments on mifepristone-induced gene regulatory systems
The mifepristone-induced gene regulatory system (M I G R S ) is a system for regulating rake gene expression through the antiprogesterone drug, mifepristone (M
Preparation of pseudotyped lentiviral vectors resistant to complement inactivation
This experimental protocol describes several methods for generating complement-stabilized lentiviral vectors either by using fusion packaging proteins containin
Lentiviral transduction of hematopoietic cells
Most hematopoietic cells, including hematopoietic stem cells and terminally differentiated cells such as primary T cells and macrophages, are undifferentiated o
Production and use of lentiviral vectors based on feline immunodeficiency virus
Lentiviral vectors based on feline immunodeficiency virus (F IV ) are very useful for mediating integrated transgenesis into non-dividing human cells. This chap
Reverse genetics of influenza viruses
It mainly describes the methods used to recover infectious influenza viruses from cDNA clones. These techniques are widely used in the integration and expressio
nuclear transplantation
Nuclear transplantation is based on the theory that all nuclei from the same embryo or cell line contain exactly the same genetic information as the fertilized
Construction of first-generation adenoviral vectors
The possibility of immune responses against proteins encoded by viral vector backbones limits the application of transgene vectors for in vivo expression. Never
Experiments with biodegradable nanoparticles
Biodegradable nanoparticles are colloidal particles, typically around IOOnm in diameter, which can be prepared from biodegradable polymers such as polylactic ac
Experiments on the theory and application of hydrophobic interaction chromatography in protein purification
Hydrophobic interaction chromatography (HIC) is a valuable tool in protein purification technology. HIC is used for protein purification on a wide range of scal
Immobilized metal affinity chromatography experiments
Summarizes the application areas of IMAC, including protein hierarchical separation and proteomics, protein fixation and detection, as well as some special appl
Down-regulation of genes generated by silencing lentiviral vectors in mice.
This chapter describes the use of lentiviral vectors to transfer genes and small interfering RNA (siR N A ) expression cassettes to preimplantation crystalline
Experiments on the isolation of subcellular organelles and subcellular structures
In proteomics research, a key step in all enrichment techniques is the subcellular fractionation, which is particularly important for the analysis of intracellu
IMViC and Hydrogen Sulfide Experiment
IMVIC is an acronym for four tests: indol test, methyl red test, Voges-Prokauer test and citrate test. i is added in English for ease of pronunciation. These fo
Stem Cell Preservation and Resuscitation
Hematopoietic stem cell transplantation is an effective treatment for malignant hematological disorders, oncological radiotherapy and severe radiation sickness
Rapid extraction of polysaccharide and polyphenol plant RNA
The reason why some plants are more difficult to extract high-quality RNA is related to a number of components enriched in these tissues, commonly phenolic comp
Source and culture experiments of human embryonal carcinoma (EC) stem cell lines
Teratocarcinomas are structurally diverse tumors that typically consist of a series of differentiated cell types that form a teratoma and a population of stem c
Generation of VSV-G-pseudotyped retroviral vectors
In order to improve the efficiency of transduction, a strategy called pseudotyping, which combines retroviral particles with other virally encoded capsid protei
Construction of multiple cis-trans vectors linked by 2A polypeptides
This chapter describes methods for expressing multiple proteins from a single readable frame by using 2A polypeptide-linked polycis-trans vectors. When these sm
MARK signaling pathway
The mitogen-activated protein kinase (MAPK) signaling pathway is a signaling pathway that is widely present in various cells and consists of a set of cascade-ac
Methylation PCR
Methylation-Specific PCR (MS-PCR) is a new method developed by Herman et al. (1996) based on the use of bisulfite treatment. This method is highly sensitive and
Apparent molecular weight determination experiment
Source: Laboratory Guide to Proteins and Proteomics
Embryonic stem cell in vitro culture and directed induction of differentiation experiments
Feeder cells are necessary for embryonic-derived stem cells including embryonic cancer cells (EC), embryonic stem cells (ES), and primordial germ cells (EG) to
Stable virus-producing cell lines for AAV assembly
Cloning of virus-producing cells by infection with a helper virus, stable virus-producing cell lines containing and genes, and r A A V (recombinant adeno-associ
Delivery of genes with HSV/AAV heterozygous amplicon vectors
This chapter describes how to construct H S V /A A V heterotrimeric vectors using an unassisted viral packaging system, and also discusses its advantages and di
Small RNA virus-based expression vectors
Small R N A viruses comprise a large group of small, pod-less, positive-sense single-stranded R N A viruses. Small R N A viruses have a rapid life cycle that ta
Production and characterization of helper virus-dependent adenoviral vectors
This chapter describes in detail the preparation, amplification and large-scale production of helper virus-dependent adenoviral vectors (H D A d ) for use in ge
In vitro packaging of SV40: a pseudoviral gene delivery system
Nuclear extracts from baculovirus-infected insect cells of the meadow moth (Sf9 ) encode the major capsid protein (V P l ) of simian virus 40 (S V 4 0 ). In the
Measurement of Ph in polluted water
Food factories require that the pH value of wastewater discharged be between 6.5 and 8.5. In the production process of fruit and vegetable sweat drink factories
Construction of siRNA expression vector targeting EGFR gene
The constructed psj-EGFR vector was transfected into A549 cells using Lipofeetamine2000, and real-time quantitative fluorescence PCR, immunofluorescence and flo
Gene transfer into mammalian cells using targeted filamentous phages
Because of their genetic simplicity, phages are commonly used in vector genetic optimization approaches for therapeutic gene transfer. In addition, since phage
Sequencing Principles and Procedures for the ABI Model 310 DNA Sequencer
DNA sequencing is divided into manual sequencing and automated sequencing, and manual sequencing includes the Sanger double deoxy chain termination method and t
One-way gel electrophoresis experiment
The S D S -- P A G E methods presented in this book will fulfill the needs of most applications in this field. Authors: Burgess et al, Translator: Chen Wei, Thi
SIV Vectors as DNA Delivery Mediators
Provides some methods on how to use the miniature SIV packaging system and SIV gene transfer vectors to obtain high titer and safe lentiviral vectors developed
Basic techniques for the preparation and use of PPA
Cross-linking of the initially purified SPA with HRP by sodium periodate oxidation (cross-linking reaction time at 22°C for 5h) gives PPA.
Simian vacuolar virus type 1 vector
Vacuolar viruses (FV) are non-pathogenic retroviruses that efficiently and safely transfer genes to different types of cells in different species. These viruses
Stem Cell and Gene Therapy
Gene therapy refers to the suppression, replacement or supplementation of defective genes at the genetic level through the regulation of the expression of targe
Feulgen staining of DNA
DNA is hydrolyzed by a weak acid (1 mol/L HCl), and the bonds between the purine bases on it and the deoxyribose sugar are opened, causing the formation of free
Experiments in the culture of human germ cell lineages
This chapter focuses on embryonic germ cells (E G C ) and aims to provide a series of experimental protocols that can be utilized to understand the relationship
Translocation of target genes with surface-modified lentiviral vectors
Two types of surface modifications have been made to lentiviral vectors in order to achieve target gene translocation: (i) taking advantage of the targeting pro
Hidradenitis elegans nematode RNA preparation
A modification of the standard method for RNA preparation from mammalian tissues and Drosophila that focuses on rapid lysis of Juniperus communis and homogeneou
Transfection of hippocampal neuronal cells with calcium phosphate co-precipitated plasmid DNA
The use of DNA/phosphorothioate (CAPO 4) co-precipitation to transfect cells is a widely described method. Although this co-precipitation method was proposed mo
Cell and Tissue Targeting
Currently, the life cycle and biology of adenovirus (A d ) infected cells have been well characterized. In A d virus infection and transgene expression, there a
Polyomavirus: SV40
This chapter describes how to construct H S V /A A V heterotrimeric vectors using an unassisted viral packaging system, and also discusses its advantages and di
Generation of foamy virus vectors and transduction of hematopoietic stem cells
Foamy virus (F V ), or spumavirus, is a nonpathogenic retrovirus developed as an integrating viral vector. Several potential advantages of F V vectors include a
X-ray crystallography for protein structure determination
X-ray diffraction is an extremely important method for determining the crystal structure of proteins. Biomolecule X-ray crystallography is the science of reveal
HIV-2 Vectors for Human Gene Therapy: Design, Construction, and Therapeutic Prospects
Lentiviral vectors can be designed as replication-incomplete or replication-condition-dependent (motile) vectors. HIV-2 vectors are more effective in gene thera
Experimental environmental metabolomics studies with 1H-NMR spectroscopy
Environmental metabolomics is a branch of metabolomics that focuses on the metabolic changes of organisms in response to environmental stress. Because this appr
Experiments to determine the presence of protein subunits and their size and molecular mass
The size or apparent atomic mass of proteins is perhaps the most common way of distinguishing the nature of a molecule. As a basis for many separation methods,
Cell-free translation experiments of integral membrane proteins in single-compartment liposomes
Membrane proteins provide the appropriate molecular mechanisms that allow the controlled entry of useful molecules into the cell and permit the export of intrac
Liquid Nitrogen Grinding
Liquid Nitrogen Milling for Applications Related to Deoxyribonucleic Acid (DNA) Isolation
Preparation of pH standard color scale
Most cell lines grow best at pH 7.4, with little variation between cell lines; however, some normal fibroblast cell lines grow best at pH 7.4 to 7.7. However, s
Transcription cDNA probe
A probe that has obtained mRNA from the transcription of the corresponding gene, which is then obtained by reverse transcription, is called a cDNa probe (cDNa p
Parallel method experiments for expression and purification
Proteins are diverse in nature, which makes it challenging to express and purify proteins using parallel methods. Parallel methods can especially be applied whe
Screening, isolation and characterization of target peptides for ligand-directed gene delivery
Functional vascular mapping of tissue-specific and angiogenesis-related receptors has been more widely promoted and optimized by isolation of receptors, cell li
Separation techniques of oxygen-sensitive proteins: [4Fe-4S]-FNR as an example
Describes the methods used to isolate the oxygen-sensitive iron-sulfur protein FNR in the laboratory. Authors: Burgess et al., Translator: Chen Wei, this experi
PCR-ELISA Telomerase Assay
Telomeres are specific DNA-protein structures at the ends of eukaryotic chromosomes, and telomeric DNA is a series of repetitive G-rich DNA sequences, which are
Protein transfection induced by ips cells
Heterodimeric expression of Oct4, Klf4, Sox2, and c-Myc transcription factors enables reprogramming of mouse somatic cells to induce functional stem cells. (1)
G protein-coupled receptor pathway: GC-cGMP-PKG
Extracellular signals are received by receptors and delivered to AC via Gs or Gi to activate or inhibit it; AC is activated to produce cAMP, which activates PKA
Targeted electrically neutral lipid vesicles for systemic delivery of genes
This chapter describes the detailed experimental procedure for encapsulation of nucleic acids by nanolipid particles (N L P ). This N L P carries a polyethylene
Experimental differentiation of embryonic stem cells to cardiomyocytes
Describes several methods used to generate and differentiate cardiomyocytes differentiated from hESC, which are as effective as the methods used to isolate card
Vesicular stomatitis virus G protein binding assay
The capsid fusion G protein of vesicular stomatitis virus (VSV-G) can be used to prepare pseudotyped retroviruses and pseudolentiviruses, and this glycoprotein
Optimizing in vitro mammalian cell electrotransfection
Electrotransfection can be used for studies of gene function, promoter activity, and gene regulation; it can also be used to create transgenic or knockout embry
Transposon-mediated delivery of small interfering RNAs: the "Sleeping Beauty" transposon experiment
A method for efficient delivery of pol III-driven SHRNA expression frames using the Sleeping Beauty (SB) transposon. Advantages of this system include its ease
Water-soluble liposomes and lipopeptides for nucleic acid delivery
Water-soluble lipopolymers and lipopeptides are a class of nonviral gene vectors that combine the increased permeability of lipid materials to cell membranes by
Naked DNA transporter assay using high-pressure injection technique
Among the various viral and non-viral means commonly used for nucleic acid transport and gene expression in animal cells, the naked plasmid D N A technique has
Cell penetrating peptide-mediated peptide nucleic acid oligomer delivery assay
Site-specific knockout and non-toxic DNA delivery technologies are particularly important in the study of protein function and as a means for future therapies.
Solid lipid nanoparticles for in vitro mammalian cell transfection
Solid lipid nanoparticles (S L N ) have many technical advantages over gene carriers such as cationic liposomes and cationic polymers. However, in the absence o
High-throughput method for screening polymer transfection reagents
The development of a safe and effective gene delivery system is a prerequisite for the success of clinical gene therapy. Despite the high transfer efficiency of
Cross-linked hyaluronic acid matrix and film experiments with specific plasmid DNAs
Natural polysaccharides are being extensively studied as carriers for gene delivery. The non-inflammatory and non-immunogenic nature of hyaluronic acid (H A ) i
PEI Nanoparticles for Targeted Gene Delivery
P E I Due to its polycationic nature, it and negatively charged D N A molecules form nanoparticles through electrostatic interactions, which further help D N A
Linear polyethyleneimine: synthesis and in vitro and in vivo transfection manipulation experiments
Linear polyethyleneimine (L P E I ) is a very effective transfection reagent suitable for large numbers of cell lines and primary cells, and it is equally suita
Intrauterine microelectroporation for efficient gene transfer in mouse embryos
The introduction of genes into the embryo has been successfully achieved by a new technique of electroporation of the chicken embryo. In contrast, since the mam
Experiments on nanogene vectors based on polylactic acid and polyethylene glycol
Nanoparticles combining polylactic acid-hydroxyglycolic acid (PLGA) and polyethylene glycol (PEG) derivatives have been used as gene carriers for mucosal permea
Experiments on photochemical internalization techniques for light-directed gene delivery
Photochemical internalization (P C I ) allows efficient transfection by a mechanism whereby light is directed to disrupt endocytosis vesicles in cells (Berg et
Preliminary enrichment experiments of Affi-Gel BLUE and nucleotide-bound proteins
While concentrating the sample, nucleic acids can be removed and the target protein can be enriched to a certain extent. We have found a remarkable resin, Affi-
Sonopuncture: an effective technique for introducing genes into chicken embryos
The acoustic pore effect may not be applicable to the introduction of genes into tissues with lumen-forming or open structures. Acoustic pore effects on neural
Neural differentiation of human EC and ES cells
The study of both E S and E C cell differentiation has facilitated the development of experimental techniques. E S cells are very sensitive to differentiation i
Efficient DNA delivery by phage into mammalian cells
Transferring genes into cells and expressing new proteins, or in this way altering the expression levels of endogenous proteins, can modulate the physiological
Chromatin open sequencing
Chromatin accessibility is also commonly understood as the degree to which chromatin is open.The replication of DNA, as well as its transcription, requires that
Genetic experiments for manipulating mammalian cells by microinjection
Introduction by microinjection can be used for any type of adherent cell, including primary cells. Since siRNA for any target gene can be obtained simply and ra
PEGylated nanoparticles of poly(L-lysine) and DNA
Nanoparticles of P E G-coated poly(L-lysine) and D N A are water-soluble, stable in saline and tissue fluids, transfect undivided cells (Liu et al. 2003), show
Reassembly experiments of human artificial chromosomes based on bacterial artificial chromosomes
Successful gene transfer requires efficient transport and long-term presence of exogenous genes in order to maintain physiologically relevant and appropriately
Experiments on a highly sensitive gene regulatory system based on a synthetic small-molecule ligand-conducted decidual hormone receptor
The RheoSwitch system is an efficient gene regulatory system based on the ecdysone receptor, which is activated by synthetic small molecule ligands. The recepto
DNA damage
DNA damage
lipoplex and LPD nanoparticles for in vivo gene delivery
The core of gene therapy research lies in the development of suitable delivery systems. Although most clinical experiments currently utilize viral carrier syste
Magnetic transfection experiment
Magnetotransfection is defined as the delivery of nucleic acids or nucleic acid carriers by magnetic particles mediated by magnetic forces. The carriers can be
Expression and characterization of ribozymes and short hairpin RNA in mammalian cells
Functional genomic research using R N A interference techniques (R N A i ) has been carried out extensively in recent years.R N A i is the newest and most succe
HVJ Liposomes and HVJ Packaging Vectors
This chapter describes the construction of a fusion-mediated vector based on inactivated H V J (hemagglutinating virus of Japan, also known as Sendaivirus). The
Cationic polysaccharide assay for DNA delivery
Polycations are an effective class of non-viral gene delivery vectors. These vectors vary in molecular mass, chemical structure, polymer/DNA ratio, and molecula
Glucose loading test
None
In vitro modeling of the blood-brain barrier
The Blood-Brain Barrier (BBB) is a highly specialized biological barrier composed of microvascular endothelial cells, basement membranes, and surrounding astroc
Molecular Interaction Detection
Isothermal Titration Calorimetry (ITC) is a commonly used biophysical experimental technique to study the interactions between biomolecules by measuring the pro
Preparation of dsRNA
Preparation of dsRNA
spotted hybridization experiment
Spot hybridization (dotblot) is a technique for detecting, analyzing, and characterizing DNA, RNA, and proteins, and determines whether or not there is hybridiz
antibody marker
None
Endolysin activity assay
As a phage-derived enzyme, endolysin is essentially a protein molecule that assists in the release of phage particles from the bacterial host by breaking down t
Measurement of superoxide dismutase (SOD) concentration
Mouse SOD was determined by enzyme-linked immunosorbent assay (ELISA). By double-antibody one-step sandwich method, in a 96-well plate coated with SOD antibody,
Subcutaneous graft tumor model construction
A certain amount of tumor cells were injected subcutaneously into nude mice with congenital immunodeficiency to form tumors, thus observing the development of t
Thermostatic Amplification Technology
Ring-mediated thermostatic amplification refers to the technique of nucleic acid amplification in a short period of time by using four specific primers at 60~65
fluorescence immunochromatography
Fluorescent Immunoassay (FIA) is an analytical technique based on antibody-antigen specific recognition to detect and quantify mobile target molecules by fluore
Blood-brain barrier permeability assay
Evans blue staining is of very wide use in in vivo experiments in small animals involving the cardiovascular system, nervous system, skeletal muscle system and
Fluorescence In Situ Hybridization (FISH)
Fluorescence in situ hybridization (FISH) is an emerging molecular cytogenetic technology, a non-radioactive in situ hybridization technique developed on the ba
genotyping
Mouse genotyping with mouse tail or toe or tissue
Animal electrophysiology experiment
None
Measurement of nitric oxide content
NO (Nitric Oxide, NO) is widely distributed in living organisms, including the nervous, circulatory, respiratory, digestive, urogenital and other systems, espec
fluorescence co-localization
None
Determination of virulence of pathogenic bacteria
Virulence indicates the strength of a pathogen's ability to cause disease, and the substances that make up bacterial virulence are called virulence factors, mai
Detection of risk zones and infarct zones for ischemia-reperfusion injury in the heart
Evans Blue staining is widely used in in vivo experiments in small animals to study diseases of the cardiovascular system, nervous system, and skeletal muscle s
Fungus Detection Laboratory
Silver hexamine staining is also known as (Periodic-acid silver methenamine) PASM staining. The specimen is oxidized by periodate, which exposes the aldehyde gr
isolated heart perfusion assay
Mouse ex vivo cardiac perfusion is a commonly used physiologic experiment to assess physiologic function and drug effects in the mouse heart. The experiment sim
In vitro reconstruction of neuronal loops: experiments on methods for building a simple model system
We can reconstruct the S-cell network that initiates the basic respiratory behavior on cultured cells of the freshwater mollusc phylum Cnidaria by using in vitr
Construction of a subcutaneous hormonal tumor model of melanoma
The B16F10 melanoma subcutaneous hormonal model is simple to operate, low cost, reproducible, and has a relatively short experimental period, making it a common
Expression purification of endolysin
As a phage-derived enzyme, endolysin is essentially a protein molecule that assists in the release of phage particles from the bacterial host by breaking down t
cell irradiation culture
Cell irradiation refers to the process of irradiating cells with ionizing rays such as X-rays, B and y-rays.
HIV drug resistance testing
HIV drug resistance testing can be phenotypic or genotypic testing, genotypic testing is the most commonly used testing method, the method is mainly through the
Amino acid content measurement experiment
The Kjeldahl method is a commonly used chemical analytical method for determining the nitrogen content of organic and inorganic samples. It was originally inven
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