Cytoplasmic RNA extraction
Since there are many sources of RNA species, there are different methods of extraction and preparation, generally phenol method, detergent method and guanidine
主题
RNA Lab
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。
107 项资源 · 实验方案、常见问题与文章
Fungal RNA extraction
Fungal RNA extraction can be: (1) used in the study of fungal RNA interference mechanism; (2) used in cDNA library construction and other studies; (3) used in t
RNA extraction and labeling experiments
None
Chromosome preparation experiments on cultured peripheral blood cells
Although chromosomes can be prepared from other cells, human peripheral blood leukocytes are the easiest to synchronize, allowing high-resolution analysis of th
Yeast Spliceosome Analysis Experiment
A precursor is a macromolecular ribonucleoprotein complex that splices mRNA precursors in the nucleus of eukaryotic cells. It consists of five snRNPs and a larg
Experiments on the isolation and analysis of pre-splitters and spliceosomes
The spliceosome is a ribonucleoprotein body (RNP) composed of RNA and proteins, which removes introns from precursor mRNA during the splicing process of precurs
In vitro splicing analysis experiments in mammalian cells
It is typical for splicing reactions to use nuclear extracts, i.e., S100 extracts complementing partially purified fractions of SR proteins or crude extracts, m
Experiments on the preparation of inhomogeneous ribonucleoprotein complexes
In the nucleus, the transcription products of RNA polymerase II bind to a large number of different types of nuclear precursor mRNA/mRNA-binding proteins, heter
Experimental purification and removal of nucleoprotein complexes by antisense affinity chromatography
This experiment was derived from "RNA Laboratory Guidebook", edited by Xiaofei Zheng.
In vivo isolation of ribonucleoprotein complex by immunoprecipitation-rPCR of RNA
The RNA Immunoprecipitation-Randomized Polymerization Chain Reaction method can be used to identify RNA components in intracellular RNP complexes. This experime
deoxyribonuclease assay
With the development of in vitro molecular experimental techniques, many DNA molecules with different catalytic functions have been discovered in recent years.
Experimental detection of protein-double-stranded RNA interactions by methylene blue-mediated photocrosslinking reaction
Methylene blue-mediated photocrosslinking reactions are a complement to UV crosslinking reactions. Methylene blue belongs to the phenothiazine group of dyes. Du
Solubilization and preservation of RNA in formamide
Storing RNA in formamide has many advantages over storing it in water or ethanol: (1) formamide will protect the RNA from degradation by RNA enzymes, and RNA st
Biotinylated oligodeoxythymidine-streptavidin magnetic bead purification band Poly(A)+RNA
The use of magnetic microspheres instead of transferring fiber cords as a medium is more commonly used and has been commercialized.
Oligodeoxythymidine cellulose purification band poly( A)+RNA
Most mammalian messenger RNAs (mRNAs) carry a poly(A)+ tail, and Poly(A)+-carrying RNAs can be isolated from total RNA using this protocol.
TRlzol Reagent One-Step RNA Isolation Assay
Animal tissues and cells are the most commonly used experimental materials in molecular biology research, a typical mammalian cell contains about 105 ug RNA, wh
Toluidine blue staining test
Toluidine blue is one of the commonly used synthetic dyes, belonging to the quinone imine dyes, which generally contain two chromophores, an amine group and a q
Hematoxylin/Eosin staining experiment
This method of staining is based on the fact that tissue structures bind differently to different dyes. The dye hematoxylin stains basophilic structures blue-vi
Giemsa staining experiment
Staining of cells allows (1) coloring of cells for easy observation and (2) detection of apoptotic cells.
In situ hybridization experiments on frozen sections
In situ hybridization of cellular RNA is used to localize specific mRNAs in mixed cell populations and tissues. Frozen section is a method of slicing tissue aft
In situ hybridization experiments on paraffin sections or cells
In situ hybridization experiments for cellular RNA are used to localize specific mRNAs in mixed cell populations and tissues.
Identification of chordoma-associated genes and screening experiments
Chordoma-associated gene identification and screening assays for (1) molecular mechanisms of chordoma (2) gene diagnosis (3) gene therapy.
Influenza virus RNA extraction
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Encephalitis B virus RNA extraction
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Hepatitis A virus RNA extraction
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Recovery of RNA fragments from polyacrylamide gels
The best method for recovering RNA fragments from poly(lactonamide) gels is the crush-and-soak method, which provides excellent recovery of high-purity single-s
RNA standard transcription reaction
The 3'-5' exonuclease activity of the large fragment of DNA polymerase I (Klenow enzyme) was utilized to convert the 3' sticky ends into flat ends. The method i
RNA bulk transcription synthesis
The typical recovery rate of the RNA standard transcription system is lugRNA/ug plasmid DNA, which can be increased to 5~l0ug RNA/ug plasmid DNA by using the fo
Photo-activated nucleotide analog-mediated RNA-protein cross-linking experiments
Photochemical cross-linking technology is an effective means to study the interaction between RNA and protein in nuclear enameled egg complexes. The basic princ
PAT method for analyzing mRNA Poly(A) tail length assay
The PAT method [ PCR poly(A) test ] can quickly and sensitively analyze specific mRNA from total RNA and quantitatively estimate the length of RNA poly(A) tails
Fe(II)-EDTA protection assay experiment
Fe(II)-EDTA is capable of catalyzing the breakage of RNA or DNA and is a complementary method to RNase protection analysis. It has the advantage that there is l
RNA in situ hybridization assay
In situ hybridization (ISH), also known as hybrid histochemistry or hybridization for cytology, is a technique that enables morphological demonstration of the p
Primer extension method RNA analysis
The primer extension method can be used for RNA mapping and RNA 5' halo. In this method, an excess of a 5' end-labeled single-stranded RNA primer is hybridized
RNA SI Nuclease Mapping
SI nuclease is an endonuclease that was isolated from Aspergillus oryzae. It can degrade single-stranded nucleic acids but not double-stranded nucleic acids. In
Spot and slit hybridization
Spot and slit hybridization analysis of RNA enables the qualitative or semi-quantitative determination of RNA by directly spotting RNA samples onto nitrocellulo
Methylmercury hydroxide agarose gel electrophoresis
Methylmercury hydroxide reacts with the imine bonds on uracil and guanine in RNA, which are involved in Watson-Crick base pairing. Therefore, methylmercury hydr
Glyoxal/DMSO denaturing agarose gel electrophoresis
Under slightly acidic conditions, the two acetaldehyde groups of glyoxal interact with the imino group of guanosine to form a cyclic compound, which inhibits th
Formaldehyde denaturing agarose gel electrophoresis
Agarose gel electrophoresis is commonly used to isolate RNA prior to Northern blotting.
RNA spectroscopy and quantification
Nucleic acids are accurately quantified using spectrophotometry because this method does not destroy the structure and also allows for sample recovery. RNA has
Extracellular system for translating viral mRNA
Translation of poliovirus and hepatitis C virus mRNA with mammalian cell S10 extracts.
A rapid response system for coupling in vitro transcription with in vitro translation
TNT 耦联网织红细胞裂解物系统乂可以分为：TNTT7 系统、TNTSP6 系统和 TN 下 PCR 系统。 The TNTT7 system translates proteins from both linear and circular DNA, the S
In vitro translation system for prokaryotic cells
The S30 family of extracts is divided into three categories: the E. coli S30 Extract System for in vitro translation of circular DNA, the E. coli S30 Extract Sy
An in vitro translation system for African Xenopus oocytes
The African Xenopus oocyte in vitro translation system is capable of sustained modification and processing of most exogenous mRNAs due to its ability to translo
Translation of mRNA in wheat germ extracts
Wheat germ extract in vitro translation system for efficient translation of mRNA in viruses and eukaryotic cells in a heterogeneous cell-free system
Translation of mRNA in reticulocyte lysates
The mRNA extracted from mammalian cells or produced by in vitro transcription of cloned DNA can be translated in cell-free extracts to synthesize proteins, whic
mRNA 3' end processing in vitro
There are two strategies for in vitro experiments; one is to co-incubate a plasmid containing the target gene at the Po ly( A) locus with cell-free extracts to
Deoxyribonuclease cleaves RNA at a specific site
Deoxyribonucleases are oligodeoxyribonucleotides consisting of about three dozen deoxyribonucleotides, which are easy to prepare and convenient to use, and are
Nuclease cleaves RNA at a specific site
Nuclease is a useful tool for obtaining site-specific cleavage products of long fragments of RNA for the study of RNA modification, for modification site analys
Ligation of RNA molecules with T4 RNA ligase
T4RNA ligases have been used to generate many site-specific modified RNAs, especially oligonucleotide-modified and tRNA-modified with antisense codon tRNAs.In a
T4 DNA ligase ligates RNA molecules
T4 DNA Ligase joins double-stranded complexes, including RNA-DNA heteroduplexes and RNA-RNA heteroduplexes, defectively.
RNA reverse transcription
The RNA template is used to obtain DNA by reverse transcription, which is then used to replicate and infect cells.
RNA radiolabeling
Intramolecular labeling of RNA during transcription, post-transcriptional labeling of the 5' end of RNA using T4 polynucleotide kinase and labeling of the 3' en
nuclear transcription analysis
Nuclear runoff (runoff) analysis can be used to examine the transcriptional efficiency of specific genes in isolated cell nuclei. For runoff analysis, the nucle
A simple and effective transcription method to reduce heterogeneity at the 3' end of RNA
Ribosyl C2'-methoxy modification of the nucleotides at the 2 5' ends. Amplification of DNA by PCR with modified primers still reduces N+l activity, and since PC
Synthesize 5' capped transcripts
Some RNA transcripts require an m7 G(5')Ppp(5')G cap at the 5' end, and they have higher translational efficiency only in cell-free extracts or in African Xenop
Effect of different methods of treating tissues on the yield of extracted RNA
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Rapid isolation of total RNA from mammalian cells
SDS lyses the cells and extracts the total RNA by acid phenol graded extraction, which is a suitable method for treating cells with relatively low mRNA content
Extraction and isolation of rat liver rRNAs
Rat liver was homogenized, the supernatant was centrifuged, and RNA was extracted with a mixture of phenol and m-cresol. rRNA and mRNA were separated in the pre
RNA extraction from formalin-fixed, paraffin-embedded tissues
Formalin-fixed, paraffin-embedded tissues from the pathology department can be used for RNA expression analysis. RNA isolated from biopsy, surgical, and cadaver
RNA extraction from fresh and frozen blood
Whole blood contains nucleated leukocytes, so it is easy to extract RNA from blood, but red blood cells are rich in ribonuclease, so it is difficult to isolate
Extraction of RNA from tissues by the CsCl method
Because some tissues (e.g., pancreas and uterus) contain high concentrations of endogenous RNAases, more care must be taken in purifying tissue-derived RNA.
Extraction of RNA from cultured cells by the CsCl method
RNA preparation by guanidine thiocyanate extraction and cesium chloride centrifugation is suitable for isolation of RNA from cells with difficult separation of
Total RNA extraction from egg and embryo cells
This method is a simple and effective way to isolate RNA from oocytes, fertilized eggs and fetal cells of frogs, sea urchins, periphyton, worms and flies.
Isolation of cytoplasmic RNA from mammalian cells
The following method is a simple and effective method for purifying cytoplasmic RNA from tissue culture cells, which appeared at an earlier stage of RNA isolati
Viral RNA Extraction
Viral RNA extraction can be (1) used for the study of RNA viral replication mechanisms, (2) used for reverse genetics studies, and (3) used for other studies in
Principles, materials and procedures for denaturing agarose gel electrophoresis of RNA
This experiment is mainly used to determine the integrity of RNA extracts.
cDNA synthesis technology
After the mRNA of the target gene has been identified by in vitro cell-free translation, the first strand of cDNA can be synthesized in the steps described in t
Construction of siRNA expression vectors
The construction of siRNA expression vectors can be applied to: (1) as a powerful tool for gene function analysis in the post-genomic era; (2) genomics and cell
Whole blood RNA extraction
Whole blood RNA extraction can be used (1) to study the mechanism of RNA interference, etc.; (2) as a template for reverse transcription; and (3) for other stud
Northern blotting and narrow line hybridization analysis of RNA
Northern blot is one of the most rigorous methods to study gene expression, which can quantitatively analyze the expression abundance of a specific mRNA in tiss
Poly(A)+ RNA Preparation Experiments
The use of cND A microarray technology can be used to (1) analyze and detect the expression of thousands of genes in parallel; (2) play an important role in the
Plant RNA preparation experiment
Since there are many sources of RNA species, there are different methods of extraction and preparation, generally phenol method, detergent method and guanidine
Preparation of fungal total RNA
Total RNA was extracted from a variety of different fungi.
Yeast precursor mRNA splicing extract assay
The splicing of eukaryotic pre-mRNA is a two-step process, in which an intron is cut from the precursor and two exons are joined together to form the mature mRN
Analysis of mRNA degradation experiments by cell-free extracts of higher eukaryotes
Changes in the environment of the cell, the state of the replication cycle, and the state of differentiation can cause changes in the half-life of some mRNAs. T
Preparation of nuclear and cytoplasmic S100 extracts of HeLa cells for in vitro splicing reaction assay
Cytosolic extracts can be used for splicing of precursor mRNAs. The S100 extract, although it includes many splicing factors, does not have splicing activity be
Purification of splicing factor SR protein assay
Precursor mRNA ( pre-mRNA ) splicing factors The SR (serine/argininc rich) family of proteins is an important player in the assembly of splices onto precursor m
Preparation of antisense RNA
RNA can also be synthesized in vitro by RNA polymerases (e.g., T3, T7, or SP6 ) using linear DNA as a template. This experiment is based on the "RNA Laboratory
Purification experiments of U series small ribonucleoprotein particles
The splicosome is the catalytic unit for excising introns from primary transcripts in eukaryotes. The splicosome contains four small nuclear ribonucleoprotein p
miRNA cloning assay
In addition to the familiar mRNA, there are also a large number of non-coding RNA (ncRNA) that do not code for proteins. Non-coding RNA plays a very important r
SELEX screening of randomized RNA libraries
SELEX screening experiments of randomized RNA libraries can be used for (1) in vitro diagnosis; (2) in vivo therapy, etc.
SELEX screening of single-stranded DNA libraries
SELEX screening experiments of single-stranded DNA libraries can be used for (1) in vitro diagnostics; (2) in vivo therapeutics, etc.
Isolation of RNA from Gram-positive bacteria
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Northern Blotting Technology
Blot hybridization is the process of binding a nucleic acid fragment to be tested to a certain solid phase support and then hybridizing it with a labeled probe
Detection of RNA-RNA binding protein interaction assay using E. coli
Binding of heterologous RNA-BPs to target RNAs in E. coli inhibits translation of target genes. This method has been successfully applied to the study of a vari
Psoralen-mediated photocrosslinking reaction to study the interaction between RNA and protein molecules experimentally
An RNA-protein molecular complex analysis method based on the photochemical reaction of psoralen enables the analysis of the topological conformation of RNA-pro
Inhibition of bacterial transcription termination assay for screening RNA-binding protein assay
A bacterial anti-transcription termination detection system for studying peptide-RNA interactions by screening recombinant cDNA libraries. This experiment was d
shRNA Design
Design Short Hair Clip RNA
Phage display technology studies RNA binding protein experiments
Phage display technology is to fuse an exogenous gene with the surface protein gene of the phage, and the corresponding exogenous protein will be expressed on t
Gel blocking assay to analyze RNA-protein interaction constant assay
The simplicity of the gel reiardadon assay (also known as mobthty shift assay) method, its sensitivity, and its ability to be quantified have made it widely use
Fluorescent RNA random primer-triggered polymerase chain reaction assay
Differential display polymerase chain reaction (PCR) can facilitate the identification of new molecular markers associated with contamination exposure in many s
Extraction and purification of RNA from polysaccharide-rich plant tissues
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Isolation of mitochondrial RNA from leaves of green leaf plants
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Total RNA or poIy(A) +RNA from Drosophila embryos
RNA Laboratory Techniques Manual (Molecular Cloning - Laboratory Guide Series)
Determination of RNA-protein dissociation constants by nitrocellulose filter membrane binding assays
The binding of proteins to DNA and RNA can be determined using nitrocellulose membrane binding, which is based on the fact that most proteins bind to nitrocellu
RNA footprinting and modification interference analysis experiments
RNA footprinting and modification interference analysis are techniques used to study RNA-protein interactions.The principles of RNA sequencing and its structura
RNaseH cleavage protection experiments for oligonucleotide targeting in RNA- protein complexes
Oligonucleotide-targeted RNase II digestion assay is a method to analyze RNA protein complexes (RNP, ribonucleoprotein particles), both for crude extracts and p
Large-scale preparation of Drosophila RNA
This method allows for the preparation of RNA from 100 Drosophila or approximately 2g of Drosophila embryos.
Analysis of RNA-peptide interactions by PACE
The polyacrylamide coelectrophoresis (PACE ) assay is a relatively recently developed method for quantitatively analyzing the interaction of proteins and peptid
Yeast three-hybrid system for detecting and analyzing RNA-protein interactions
The yeast three-hybrid system can be used to analyze protein-RNA interactions. In the yeast three-hybrid system, RNA-protein interactions lead to transcription
Liquid-phase screening of RNA-binding proteins in cDNA expression libraries
The method of immobilizing cDNA library-expressed proteins on nitrocellulose or nylon membranes and then screening them with labeled RNA is the most commonly us
Northwestern screening of protein expression library assay
Northwestern screening of protein expression libraries is performed by constructing IPTG-inducible fusion protein expression libraries (the N-terminus of the fu
Natural RNA library screening and protein-specific binding RNA sequence experiments
In cells, protein-RNA binding is involved in a variety of physiological regulation and developmental processes. Direct protein-RNA interactions occur during mRN
Small-scale rapid preparation of Drosophila RNA
Guanidine hydrochloride rapidly inhibits RNAase activity while lysing cells, and this method utilizes this feature to isolate Drosophila RNA.
Preparation of yeast total RNA
Good quality total RNA can be extracted from yeast cells.
Preparation of yeast tRNA by phenol method
By purification on DEAE cellulose columns, small amounts of DNA, large molecular RNA, proteins, polysaccharides and other impurities were removed, and finally a
RNA interference technology
Post-transcriptional gene silencing (PTGS) is known as RNA interference (RNAi), whereby double-stranded RNA (dsRNA) consisting of positive-sense and negative-se
相关主题
常与免疫学实验一起研究的技术。
危险品化学品经营许可证（带存储）