multiplex PCR
Multiplex polymerase chain reaction (MPCR), also known as compound PCR, is a new PCR amplification technique improved and developed on the basis of conventional
主题
PCR technology
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。
128 项资源 · 实验方案、常见问题与文章
PCR-SSCP technology experiment
Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) technology can be used for: (1) screening and detection of oncogenes and oncogenic
Restriction fragment length polymorphism (RFLP) analysis
Restriction fragment length polymorphism (RFLP) is one of the most important analytical methods in molecular biology and is mainly used to (1) detect DNA sequen
Primer extension experiment
Primer extension analysis was used to determine the location and quantify the amount of the 5'-end of a specific RNA. To end the labeled oligonucleotide hybridi
Experimental purification of PCR products with dUTP
The incorporation of dUTP into the replicon during PCR amplification is one of the most efficient and widely used methods for purifying PCR products (Longoetal.
PCR amplification of high GC content fragments
The following program describes in detail the PCR amplification of high GC content antique fragments. This experiment was derived from PCR Laboratory Guide (Sec
Precision amplification experiments for large fragments
Amplify a ~5kb fragment with 10 different primer pairs, the template DNA is the same, the reaction system is 50ul, and the control has no template and contains
PCR product purification assay
A method is described for the purification of PCR products using an AmiconMicrocon-PCR filter (Millipore) that efficiently removes redundant dNTP and primers. T
PCR product quantification
Accurate quantification of the template prior to sequencing is critical. Depending on the amount of DNA used, several different methods are available. Two other
Sequencing reaction experiments
To get the right signal intensity, adjust the amount of DNA according to the size of the DNA fragments. For ABI PRISM Big Dye Terminator Ready Reaction mix (App
Purification of sequencing reaction products
Purification of the sequencing reaction product prior to electrophoresis is essential to remove excess, unbound dye terminators. Centrifugation columns can be a
Genomic DNA purification from whole blood Experimental methods
The Wizard Genomic DNA Purification Kit for isolation of genomic DNA from whole blood (10 ml) is based on a four-step procedure. The first step of the purificat
Genomic DNA isolation experiment based on silica membrane
Silica chemistry has become one of the methods for purifying high-quality genomic DNA from samples such as mouse tail clippings, animal tissues, and tissue cult
An experimental method for the isolation of genomic DNA based on magnetic beads
The Wizard Magne Sil Genomic DNA Purification System uses a solid-phase paramagnetic silica pellet to purify genomic DNA, replacing vacuum filtration and centri
Purification of PCR fragments from agarose gels
The Wizard SV Gel and PCR Purification System is a method for the direct purification of PCR fragments from agarose gels or from PCR amplification reaction mixt
Silica membrane-based purification of PCR products
The Wizard SV96 PCR Purification System provides a membrane-based method for high-yield PCR purification from 96-well plates. This experiment was derived from P
Magnetic particle-based purification of PCR products
MagneSil is a paramagnetic silica particle that acts as a mobile solid-phase material for capturing double-stranded DNA fragments. The target complexes adhering
Clean purification experiments for stained ends
Automated fluorescence sequencing using ABIPRISMRigDye end chemistry is one of the methods for efficient DN eight sequence determination. Sequence gradients are
Preparation of RNA by ultracentrifugation with guanidine isothiocyanate/cesium chloride
This method is derived from a modification of the protocol developed by Chirgwin et al. (1979). The reagents specified for Chirgwin et al. are commonly used in
RNA preparation from paraffin-embedded fixed tissues
This protocol was published by Godfrey et al. as an improvement on Fisher's protocol. Previously, protocols for purifying RNA from fixed tissues have been publi
RNA purification experiments with Trizol
The production of phenol-based reagents for RNA isolation is based on Chomczynski protocols. These protocols are most useful when obtaining RNA from RNAase-rich
Preparation of RNA from plant tissues using Trizol
The following protocol is a modified version of the Trizol protocol for RNA isolation from plant tissues in which a high-salt isopropyl fermentation precipitati
RNA purification experiments from plant tissues using ConcertPlant reagents
Outlined below is the protocol that comes with ConcertPlant Reagent (Invitrogen). The protocol is not phenol-based, but requires the addition of chloroform. Thi
Purification of total RNA using the Marligen Total RNA Rapid Purification System
The Rapid Total RNA System by Marligen Biosciences extracts RNA from a variety of samples with very consistent yield and quality. This experiment was derived fr
mRNA isolation assay utilizing Ambion's MicroPoly(A)Pure kit
This experiment utilizes Ambion's MicroPoly(A)Pure kit to isolate mRNA. This experiment was derived from PCR Laboratory Guide (2nd edition) by Seed Kang and Qu
DNA contamination removal assay after elution
The following protocol is a modification of the Dilworth and Huang protocol (DilworthandMcCarey1992;Huangetal.2000). It can be used to remove DNA contamination
On-column removal assay for DNA contamination
This protocol can be used for RNA isolation strategies using silica-based RNA binding columns, such as protocol 6, however, 100% DNA removal may not be achieved
Experiments in the preparation of frozen tissues and sections
For better results, fresh tissue should be used, because samples stored at -80°C for more than 24 h will have much lower RNA yield and integrity. Although seve
HE staining of slices for LCM
Prior to LCM, cut tissue samples are fixed, stained, and dehydrated if necessary. Authors use classical HE staining for visceral and lymph node tissues. Once th
LCM Experiment
LCM experiments have applications for (1) pathologic analysis of sections (2) analysis of lesion extent (3) clinical sample collection and reference.
RNA extraction experiments from a limited number of cells
RNA samples can be stored at -80°C for up to 6 months. Total cellular RNA is extracted using the StratageneRNA Extraction Kit, following the instructions. Alte
Single-stranded cDNA synthesis
This protocol utilizes total RNA because if 18SrRNA is used as an internal control, RNA with poly(A) cannot be used in subsequent experiments. and the RNA must
Relative RT-PCR: Determination of Primer-to-Competitor Ratio Experiments
The purpose of this scheme is to select an appropriate ratio between the 18SrRNA primer and the competitor that will mimic the expression of the target transcri
Relative RT-PCR: Sample quantification assay
Knowing the linear range of cycling parameters and the ratio of 18SrRNA primer to competitor, one can use one's own samples for relative quantitative RT-PCR.The
Competitive RT-PCR: Competitor RNA construction assay
Part I: Design of competitor; Part II: Synthesis, purification and quantification of competitor RNA. This experiment is from PCR Laboratory Guide (Second Editio
Competitive RT-PCR: Evaluation of copy number
In the preexperiment, reverse transcription should be done separately for the sample and the competitor, and then the unchanged amount of this sample reverse tr
Parameter experiments for real-time PCR
LUX primers are labeled with FAM (6-carboxyfluorescein) or JOE (6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein). In reactions without a specific template,
Establishment of a real-time PCR quantitative analysis experiment
The process of establishing a quantitative real-time PCR assay for a new target molecule involves: (i) a design step for the selection of PCR primers and probes
Real-time PCR experiment
This method utilizes the PlatinumQuantitativePCRSuperMix-UDG. When performing real-time PCR, there should be a set of equipment that can detect the fluorescence
Multivariate real-time PCR experiment
In multiplex real-time PCR, pairs of primers with different markers are added to a single reaction tube, allowing simultaneous analysis of several different gen
Reverse transcription and DNA amplification experiments with activated heat-stable DNA polymerase
This experiment introduces activated heat-stable DNA polymerase for reverse transcription and DNA amplification. This experiment was derived from PCR Laboratory
High-quality DNA purification assay
The application of a modified version of the salt precipitation method of Miller et al. (1988) for DNA purification, which does not use phenol or chloroform, is
Primer design and end-labeling experiments
Systematic analysis of SSCP parsing capabilities and limitations revealed a significant correlation between sensitivity and fragment length. The highest detecti
Preparation of template DNA, organization of experiments and PCR amplification experiments
Genomic DNA can be isolated by a variety of methods, depending primarily on the type (blood or tissue) and amount of initial material. All prepared DNA should b
Gel electrophoresis experiment
When optimizing the experimental conditions, the PCR products should be separated on gels containing glycerol and without glycerol at the same time and the resu
Reverse transcriptase in situ PCR assay
This section describes an abbreviated method for in situ PCR, specifically reverse transcriptase in situ PCR. Each of these experimental procedures is discussed
Large-scale PCR for cDNA microarrays
This protocol describes the experimental procedure to make cDNA microarrays by PCR amplification of products. This experiment was derived from PCR Lab Guide (Se
Hybridization and detection experiments with the HC ExpressArray Kit
This experiment describes the method of hybridization and detection using HC ExpressArray Kit. This experiment was derived from PCR Laboratory Guide (Second Edi
Experiments for the preparation of ablated cDNA or genomic DNA libraries
A total of 2ug of genomic DNA or cDNA is required for a single bi-directional ablation.Most methods for isolating RNA and genomic DNA are suitable for this expe
subtractive hybridization experiment
The experiment introduces subtractive hybridization, which is derived from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu Lijia.
PCR amplification of differential DNA
In the reactions described in this protocol, differential DNA is selectively amplified. There should be at least 4 reactions per experiment: 1) ablated test DNA
Mirror direction selection experiment
The major disadvantage of SSH is the presence of background clones representing non-differentially expressed DNA species in the ablated library. In some cases,
PCR-based DNA blotting experiments
For high-throughput screening, it is recommended to use 96- or 384-well PCR plates available from several thermal cycler manufacturers, or to use individual tub
Differential hybridization experiments with detection and drive DNA probes
The following 4 probes are used for differential screening hybridization. 1. detector-specific ablation probe (forward ablation probe). 2. driver-specific ablat
Genomic DNA removal experiment from total RNA
For FDD gene expression analysis, as well as the use of any other RNA-based gene expression technique, contaminating genomic DNA must be removed prior to revers
Reverse transcription synthesis of single-stranded cDNA experiment
The volume of a cDNA synthesis reaction is dependent on the number of anchored-anywhere primer combinations used. The protocol provided below, applies to 24 pri
FDD-PCR experiment
This protocol is designed as 24 PCR reactions using 3 cDNA subpopulations (RT reactions utilizing the H-T11M primer from protocol 3) with primers that are a com
Re-amplification, cloning and sequencing experiments of differentially expressed cDNAs
After cutting the latent differentially expressed cDNA from the acrylamide gel, the cDNA is re-amplified with the same anchored-any primer combination under the
Northern blot analysis assay
To confirm that the selected cDNAs are indeed differentially expressed, it is recommended that Northern blot analysis be used instead of other confirmatory tech
PCR-based DNA library screening experimental methods
This experiment introduces the PCR-based DNA library screening method. This experiment is derived from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu
Amplification of 3'-end cDNA by classical RACE
This experiment describes the amplification of 3'-end cDNA by classical RACE.PCR Lab Guide, Section 25.4, Protocol 1
Experiment to amplify the 5' end of cDNA by classical RACE
This protocol is divided into three phases. Phase 1: reverse transcription to generate the cDNA template; phase 2: addition of poly(A) tail to the end of the fi
Preparation of double-stranded DNA libraries encoding randomized peptides by PCR
PCR serves two important purposes when constructing libraries using synthetic oligonucleotides. First, by amplifying the library with primers that anneal to spe
The 5' end of cDNA was amplified by the new RACE method.
This experimental program is divided into 6 stages. Stage 1: dephosphorylation of degraded RNA; Stage 2: uncapping of intact RNA; Stage 3: preparation of RNA ol
Hat Conversion RACE Experiment
The method presented here uses a biotinylated primer to help purify the first strand product. These purification techniques are interchangeable. Similarly, usin
Analyzing substrates and products of ligation reactions by PCR
Synthetic DNA libraries generated by PCR can be digested enzymatically and ligated into a linearized expression vector system. In the scheme presented here, the
Quality control experiments of a randomized peptide DNA library by PCR
In this scheme, a random polypeptide library encoded by a plasmid vector is analyzed by PCR. The library is obtained by transforming bacteria with the product o
Quality control of genomic DNA libraries using PCR
In this scenario, human genomic DNA is treated with a mixture of restriction endonucleases (the recognition site of the endonuclease mixture is 4bp) to produce
Preparation experiments for cloning vectors
Many vectors and their derivatives have been developed for cloning PCR products, including the typical pBluescript-like vectors. These include the typical pBlue
Preparation of insertion fragment experiments
This experiment describes the preparation of insertion fragments. This experiment was derived from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu Lij
Preparation of cloning experiments
This experiment describes the method of PCR product preparation for cloning. This experiment is derived from PCR Laboratory Guide (Second Edition) by Seed Kang
Colony PCR analysis of cloned recombinants experiments
The colony PCR analysis of cloned recombinants assay can be applied to (1) analyze whether colonies produced from transformed cells contain recombinant insert f
ribose cloning experiment
This scheme is just one way of treating PCR products with RNAase. There are many alternative and effective ways of purifying the PCR product to remove the prime
Purification experiments with paramagnetic streptavidin anti-biotin protein beads
This experiment describes the purification of Streptomyces paramagneticus anti-biotin protein beads. This experiment was derived from PCR Laboratory Guide (2nd
Mutagenesis PCR experiment
The greatest advantage of mutagenic PCR is that various types of mutations are introduced in a preference-free manner, rather than obtaining a high level of sin
Rapid PCR targeted mutation assay
The key features of this PCR-SDM method include the following: 1. the concentration of template used is increased, which reduces the number of cycles and theref
PCR mutation experiments by overlap extension and gene SOEing
This experiment describes about the process of PCR mutation by overlap extension and gene SOEing. This experiment is from PCR Lab Guide (2nd edition) by Seed Ka
Codon design experiment
To date, genes have only been used for expression in bacteria, so it is not known whether this simple strategy could also work for transgenesis in higher plants
Customized 40bp oligonucleotide experiments
This experiment describes the process of how to customize a 40bp oligonucleotide. This experiment was derived from PCR Lab Guide (Second Edition) by Seed Kang a
Gel purification experiments
This experiment describes the method of gel purification. This experiment was derived from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu Lijia.
PCR assembly reaction
This experiment describes about the process of PCR assembly reaction. This experiment is from PCR Laboratory Guide (Second Edition) by Seed Kang and Qu Lijia.
Experiments for the determination of the affinity of the insulin receptor for insulin
This experiment describes the method of determining the affinity of insulin receptor for insulin. This experiment is from the Experimental Guide for Protein Pur
Join the joint experiment
The junction is constructed by allowing the two complementary primers to anneal, and the 5' ends of primers 1B and 2B should first be phosphorylated. Modern Neu
Labeling enzyme digestion to release labeling experiments
is complementary to the labeling enzyme technique in differential display (DD) experiments Modern Neuroscience Research Techniques Author(s): U. Windhorst &
Patch Flat Labeling Experiment
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
Connections for dual-label experiments
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
PCR amplification of double-labeled signatures
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
Separation experiments with double labels
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
Serialization experiments
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
clonal tandem assay
It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis
Screening of PK-120 gene by PCR method related to the ends of partially disassembled polypeptides
Partially dissecting the amino acid sequence of the peptide by about 20 residues, the PCR reaction with PCR primers encoding the ends of the peptide enables the
PCR differential display assay for mRNA
A DNA sequence corresponding to mRNA that can be recovered, cloned, sequenced, and can be used as a probe for hybridization or screening libraries. Source: Comp
TA cloning of PCR products
The TA Cloning System can be used for rapid, one-step insertion of PCR products directly into the multiple cloning site (MCS) of plasmid vectors.
Principles, materials and procedures for PCR amplification and isolation of target DNA fragments
PCR (Polymerase Chain Reaction) was discovered in 1986 by Kallis Mullis. This technology has been widely used in various fields of molecular biology, not only f
Hepatitis B Virus (HBV) by PCR
Hepatitis B virus (HBV) detection by PCR is mainly used for the diagnosis of viral hepatitis, efficacy observation and prevention research work.
PCR-SSP analysis experiment
PCR sequence-specific primer analysis can be used to: (1) detect sequence polymorphisms by PCR amplification of primers that specifically recognize a particular
Molecular Markers - AFLP Principles and Procedures
AFLP can be used for (1) construction of genetic linkage maps; (2) rapid identification of molecular markers closely linked to target genes using AFLP; (3) AFLP
Dual PCR experiment
With the large-scale cultivation of genetically modified (GM) crops, the safety of GM crops has received increasing attention, and in 2002, China required the l
Detection of soybean transgenes by dual PCR-capillary electrophoresis
Dual PCR-capillary electrophoresis for detection of soybean transgenes can be applied for rapid detection of glyphosate-resistant transgenic soybeans.
immuno-PCR
Immuno PCR (Im-PCR) is a technique for the detection of trace antigens that utilizes the specificity of the antigen-antibody reaction and the high sensitivity o
Hybridization and detection experiments of ISPCR amplified products
Polymerase Chain Reaction, abbreviated as PCR (English full name: Polymerase Chain Reaction) (also known as: Polymerase Chain Reaction), Polymerase Chain Reacti
Reverse transcription-polymerase chain reaction
PCR refers to the process of replicating a daughter strand of DNA complementary to the template DNA sequence in vitro through a three-step cycle of denaturation
Real-time quantitative PCR experiment
Real-time quantitative PCR (qPCR) was first introduced by Applied Biosystems in 1996. Conventional PCR methods allow for exponential amplification of specific D
Cloning PCR experiments that do not depend on the ligation reaction
Nowadays, PCR has become a powerful research tool in the field of molecular biology and is used in all aspects of modern molecular biology. A variety of PCR met
Fusion PCR experiment
Fusion PCR is the process of joining two DNA sequences together in an adjacent position by PCR. Generally speaking, the primer sequence designed for fusion PCR
Colony PCR experiment
Conventional PCR amplification requires multiple steps of bacterial culture and plasmid preparation before gene amplification, which is cumbersome and time-cons
Hot-start PCR experiments using heat-activated primers
PCR is widely used in genetic testing, paternity testing, blood screening, clinical diagnosis, and is also a very useful tool in molecular biology research beca
digital PCR
Digital PCR (digital PCR) is to convert the exponential multiplication signal of traditional PCR into a linear digital signal, which can be read by specific ins
PCR-single-stranded conformational polymorphisms
PCR-single-strand conformation polymorphism is a technique based on this DNA single-strand conformation property combined with gel electrophoresis to detect gen
single-cell PCR
Single-cell PCR utilizes flow cytometry to isolate a specific type of cell. Using a single cell, PCR can be analyzed at the DNA or mRNA level to find molecular
Single-molecule PCR
Single-molecule PCR, also known as homo-primer PCR, is a PCR technique in which a single DNA molecule is used as a template and the number of cycles is unlimite
Allele-specific PCR
Allele-specific PCR, also known as mismatch PCR, amplification refractory mutation system (ARMS), and mismatch amplification mutation assay (MAMA), is an altern
Simple allele discrimination PCR
Simple allel-discriminating PCR, which is based on the principle of amplifi-cation refractory mutation system (ARMS), in which primers are designed to introduce
Allele-specific PCR with L-DNA labeling
L-DNA-labeled allele-specific PCR combines allele-specific PCR with L-DNA-labeled PCR.L-DNA-labeled PCR amplifies DNA with a sequence-determined L-DNA tag.L-DNA
Cold denaturation co-amplification PCR
Cold denaturing co-amplification PCR is a method for selectively amplifying a small number of alleles from a mixture of wild-type and mutant-containing sequence
Homologous one-step fluorescent allele-specific PCR
Homologous one-step fluorescent allele-specific PCR utilizes water-soluble conjugated polyelectrolytes (CPs) containing a large number of absorbing units, and t
PCR-Denaturing Gradient Gel Electrophoresis
PCR-denaturing gradient gel electrophoresis is the process of amplifying the target gene through PCR by adding a nucleotide chain of 30-50 GC bases to the 5' en
Reverse transcription-polymerase chain reaction
Reverse transcription-polymerase chain reaction (RT-PCR) has improved the sensitivity of RNA detection by several orders of magnitude, making it possible to ana
In situ PCR
In situ PCR (in situ PCR) is a PCR reaction performed in tissue cells, which combines the advantages of in situ hybridization with cellular localization ability
Quantitative PCR
Quantitative PCR is a technique to quantify the amount of PCR starting template by analyzing the PCR end product or monitoring the PCR process using a standard
Methylation-specific PCR
Methylation-specific PCR (MS-PCR) is a site-specific methylation detection technique, which is one of the most commonly used and highly accurate methods to stud
PCR sequencing experiment
Since the introduction of dideoxynucleotide triphosphate (ddNTP) as a chain terminator by Sanger et al. (1975), the technology of DNA sequencing has developed r
Emulsion PCR experiment
In 2005, Margulies et al. published an article in Nature describing a fast and simple sequencing method based on an emulsion system for DNA amplification and a
PCR-based large fragment DNA synthesis assay
Chemical synthesis of DNA sequences provides a powerful tool for high level expression and functional studies of genes in heterologous systems. In recent years,
mutagenicity test
not available
Quantitative methylation-specific PCR
Quantitative methylation specific PCR (qMSP) is a molecular biology method for detecting the degree of DNA methylation, which can determine the percentage of DN
Real-time fluorescence quantitative PCR-FP
Real-time fluorescence quantitative PCR-FP
Polymerase Chain Reaction (PCR)
PCR technology can be used to (1) test whether infectious diseases are in a recessive or subclinical state; (2) effectively detect mutations in oncogenes and ac
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）