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3' Rapid Amplification of cDNA Ends (RACE) Protocol - http://www.nottingham.ac.uk/~mbzspd/methods/3RACE_PCR.html

Category: RNA Protocols: 3' RACE Rapid Amplification of cDNA Ends Protocols

PCR cycle steps, protocol using SuperScript II reverse transcriptase (Life Technologies) and Taq polymerase, Gene specific primer, T17 Adapter primer and an Adapter primer. Dr.Dawson, Nottingham. - [Read 3' Rapid Amplification of cDNA Ends (RACE) Protocol]

A protocol for application of Polymerase Chain Reaction (PCR) in situ hybridization for detection of - http://www.im.microbios.org/03setember98/08%20Sterflinger.pdf

Category: PCR Protocols: In-situ PCR Protocols

A protocol for application of Polymerase Chain Reaction (PCR) in situ. hybridization for the detection of hyphomycetes. Sterflinger et al 1998. - [Read A protocol for application of Polymerase Chain Reaction (PCR) in situ hybridization for detection of]

AFLP genotyping and fingerprinting Review PDF - http://taxonomy.zoology.gla.ac.uk/~rdmp1c/teaching/L4/Evolution/Session5/Mueller.pdf

Category: PCR Protocols: AFLP PCR

Ulrich G. Mueller and L. LaReesa Wolfenbarger. Amplified fragment length polymorphisms (AFLPs) are polymerase chain reaction (PCR)-based markers for the rapid screening of genetic diversity. AFLP. TREE October 1999 - [Read AFLP genotyping and fingerprinting Review PDF]

Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3837

Category: PCR Protocols: cDNA Synthesis Protocols

An oligodeoxynucleotide primer hybridized to mRNA is extended by an RNA-dependent DNA polymerase to create a cDNA copy that can be amplified by PCR. Depending on the purpose of the experiment, the primer for first-strand cDNA synthesis can be specifically designed to hybridize to a particular target gene, or a general primer such as oligo(dT) can be used to prime cDNA synthesis from essentially all mammalian mRNAs - [Read Amplification of cDNA Generated by Reverse Transcription of mRNA Protocol]

Amplification of RNA: High-Temperature Reverse Transcription and DNA Amplification with a Magnesium - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4115

Category: PCR Protocols: Reverse Transcriptase RT-PCR Protocols

Protocol uses a single thermostable RNA polymerase to perform high-specificity RT-PCR. A high-temperature RT reaction is followed by PCR amplification of the cDNA using a single thermostable poymerase, the GeneAmp AccRT RNA PCR enzyme from Applied Biosystems. The high temperature of the RT reaction enhances the specificity of primer binding and also reduces secondary structure in the template, thereby increasing the efficiency of polymerization. - [Read Amplification of RNA: High-Temperature Reverse Transcription and DNA Amplification with a Magnesium]

An Assay for RNA-Dependent RNA Polymerase Activity Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4322

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol provides a quick way to assay for RNA-dependent RNA polymerase (RdRP) activity in crude protein samples. RdRP transcription products remain at the origin during paper chromatography. - [Read An Assay for RNA-Dependent RNA Polymerase Activity Protocol]

Bidirectional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4139

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for bidirectional, blunt-end cloning of DNA fragments. The target DNA is PCR amplified and 3'-extensions are polished with Pfu DNA polymerase. The amplicon is ligated to a blunt-ended plasmid DNA, and the products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Bidirectional Cloning of PCR Products Protocol]

Construction of cDNA Libraries Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3560

Category: DNA Protocols: cDNA Library Protocols

Here, the DNA-RNA hybrids synthesized in Stage 1 are converted into full-length double-stranded cDNAs. The primers for synthesis of second-strand cDNA are created by RNase H, which introduces nicks into the RNA moiety of the cDNA-mRNA hybrids. E. coli DNA polymerase I extends the newly created 3'-hydroxyl termini, using the first-strand cDNA as a template. - [Read Construction of cDNA Libraries Protocol]

Determination of Tissue Fungal Burden Utilizing Quantitative Real Time Polymerase Chain Reaction - http://www.sacmm.org/pdf/Determination%20of%20Tissue%20Fungal%20Burden%20utilizing%20Quantitative%20Real%20Time%20PCR.pdf

Category: Fungi Protocols: Fungi Genetics Protocols

Standard operating procedure for the determination of tissue fungal burden utilizing quantitative real time polymerase chain reaction (QPCR). This standard operating procedure will provide information on how to assess fungal tissue burden of infected animals by use of a single copy (FKS) or multicopy gene (18s RNA) to assess the number of fungal cell nuclei present. - [Read Determination of Tissue Fungal Burden Utilizing Quantitative Real Time Polymerase Chain Reaction]

Dideoxy-mediated Sequencing of DNA Using Taq DNA Polymerase Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3803

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Because sequencing reactions catalyzed by thermostable DNA polymerasessuch as Taq are carried out at elevated temperatures, problemscaused by mismatched annealing of primers or templates richin secondary structure are greatly alleviated. - [Read Dideoxy-mediated Sequencing of DNA Using Taq DNA Polymerase Protocol]

Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3801

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol describes DNA-sequencing of single-stranded DNAtemplates in reactions catalyzed by Sequenase. - [Read Dideoxy-mediated Sequencing Reactions Using Bacteriophage T7 DNA Polymerase (Sequenase) Protocol]

Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3802

Category: DNA Protocols: DNA Sequencing Protocols: Dideoxy Mediated Sequencing Protocols

Protocol for dideoxy-mediated sequencing reactions using the Klenow fragment of E. coli DNA polymerase I and single-stranded DNA templates. - [Read Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase]

Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase I - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3802

Category: E Coli Protocols: E coli Nucleic Acid Protocols

Protocol for dideoxy-mediated sequencing reactions using the Klenow Fragment of E. coli DNA polymerase I and single-stranded DNA templates. - [Read Dideoxy-mediated Sequencing Reactions Using the Klenow Fragment of E. coli DNA Polymerase I]

Directional Cloning of PCR Products Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4140

Category: PCR Protocols: PCR Cloning Protocols

Protocol is for directional blunt-end cloning of DNA fragments. The target DNA is PCR-amplified, 3'-extensions are polished with Pfu DNA polymerase, and the amplicon is ligated to a blunt-ended plasmid DNA. The products of the ligation reaction are used to transform competent Escherichia coli. A restriction enzyme is added to the ligation reaction to relinearize any self-religating vector DNA. - [Read Directional Cloning of PCR Products Protocol]

DNA Sequencing Methods - http://www.genome.ou.edu/protocol_book/protocol_partIV.html#IV.C

Category: DNA Protocols: DNA Sequencing Protocols

Methods for DNA sequencing. Includes: Bst-catalyzed radiolabeled DNA sequencing; Radiolabeled sequencing gel preparation, loading, and electrophoresis; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye primers; Taq-polymerase catalyzed cycle sequencing using fluorescent-labeled dye terminator reactions; Sequenase[TM] catalyzed sequencing with dye-labeled terminators; etc. - [Read DNA Sequencing Methods]

Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol - https://commerce.invitrogen.com/index.cfm?fuseaction=iProtocol.unitSectionTree&treeNodeId=3DD21299A00419DEBAFDBC78E47CBE79

Category: PCR Protocols: PCR Optimization

Method for amplifying DNA enzymatically by the polymerase chain reaction (PCR), including procedures to quickly determine conditions for successful amplification of the sequence and primer sets of interest, and to optimize for specificity, sensitivity, and yield. The first step of PCR simply entails mixing template DNA, two appropriate oligonucleotide primers, Taq or other thermostable DNA polymerases, deoxyribonucleoside triphosphates (dNTPs), and a buffer. - [Read Enzymatic Amplification of DNA by PCR: Standard Procedures and Optimization Protocol]

Fermentas Taq DNA Polymerase Protocol - http://www.fermentas.com/techinfo/pcr/dnaamplprotocol.htm

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

Fermentas Taq DNA Polymerase Protocol. Fermentas. - [Read Fermentas Taq DNA Polymerase Protocol]

Fermentas Taq DNA Polymerase Protocol - http://www.fermentas.com/techinfo/pcr/dnaamplprotocol.htm

Category: PCR Protocols

Fermentas Taq DNA Polymerase Protocol. Fermentas. - [Read Fermentas Taq DNA Polymerase Protocol]

Home-made Taq Polymerase Purification - http://www.nottingham.ac.uk/~mbzspd/methods/Home_made_Taq_polyme.html

Category: PCR Protocols

PCR polymerase costs can be high. If you are willing to work, you can produce bacteria containing the clone. It appears to produce lots of Taq and is quite stable. The proceedure takes 4 days start to (15 000 units of Taq) finish. The Taq also appears ver - [Read Home-made Taq Polymerase Purification]

In Situ Polymerase Chain Reaction Roche - http://www.roche-applied-science.com/PROD_INF/MANUALS/pcr_man/Chapter10/CHAP10-Seite240.htm

Category: PCR Protocols: In-situ PCR Protocols

In Situ Polymerase Chain Reaction: Past, Present, Future. Roche. - [Read In Situ Polymerase Chain Reaction Roche]

In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3813

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for in vitro mutagenesis using double-stranded DNA templates. Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA. - [Read In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI]

In Vitro Transcription with SP6 Polymerase Protocol - http://www.scienceboard.net/resources/protocols.asp?action=article&protocol_id=92

Category: Cell Biology and Cell Culture: In Vitro Reconstitution Protocols

Protocol for in vitro transcription with SP6 polymerase protocol. - [Read In Vitro Transcription with SP6 Polymerase Protocol]

Long-PCR Reagents and Guidelines - http://wheat.pw.usda.gov/~lazo/methods/lazo/longpcr.html

Category: PCR Protocols: Quantitative PCR Protocols

General guidelines for long-PCR conditions and enzyme mixtures. Efficient long-PCR results from the use of two polymerases: a non-proofreading polymerase is the main polymerase in the reaction, and a proofreading polymerase (3' to 5' exo) is present at a lower concentration. Includes: For PCR with low-complexity templates (e.g., plasmid and cosmid inserts); For PCR with moderate-complexity templates (e.g., bacterial genomic DNA); For PCR with high-complexity templates (e.g., human genomic DNA). - [Read Long-PCR Reagents and Guidelines]

Method to Analyze Meiotic Segregation Patterns in Coprinus cinereus Using PCR - http://www.fgsc.net/fgn/freedman.html

Category: Fungi Protocols: Fungi Genetics Protocols

Quick and reliable method to analyze meiotic segregation patterns in Coprinus cinereus using the polymerase chain reaction. The advantages of this method include: 1. The tissue is grown and lyophilized in the same tube, which facilitates the simultaneous analysis of many segregants. 2. Only one extraction step is necessary. 3. The markers are scored by gel electrophoresis, thereby bypassing Southern analysis. - [Read Method to Analyze Meiotic Segregation Patterns in Coprinus cinereus Using PCR]

Older RT-PCR and PCR Techniques Page. - http://www.cas.psu.edu/docs/CASDEPT/VET/jackvh/jvhpcr.html#V

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

Quantitative Reverse Transcription Polymerase Chain Reaction (RT-PCR) and Other PCR Procedures. - [Read Older RT-PCR and PCR Techniques Page.]

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3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol

3' Rapid Amplification of cDNA Ends RACE Using PCR Protocol. This protocol contains the steps for 3' end rapid amplification of mRNA by PCR. The first-strand cDNA is synthesized from total or poly(A+) RNA by priming from the poly-A tail of the mRNA using a oligo (dT) adaptor primer. The cDNA is then amplified via PCR using a gene-specific primer and an adaptor primer.

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