PCR Protocols

A Magnetic Particle-Based Method for Purifying PCR Products from Solution Protocol The MagneSil system can selectively isolate PCR products that are more than 150-bp long from primers and primer -dimers.  The technology can be used with a number of robotic workstations, including Beckman Coulter’s Biomek 2000 and FX Laboratory Automation Workstations.  The procedure can also be carried out manually.  Typical recovery is more than 80% for a 1-kb product with negligible carryover of primers or nucleotides.

Basic Ideas in PCR Protocol Design. PDF Basic Ideas in PCR Protocol Design. PDF

Basic PCR Protocol Basic PCR Protocol. (per Adam 08/12/04).

Collection of Specimens for Detection of Measles Virus RNA by PCR Protocol Protocol for collection of specimens for detection of measles virus RNA by PCR.

Dealing with Carryover Contamination in PCR: An Enzymatic Strategy Protocol Protocol for dealing with carryover contamination in PCR- enzymatic strategy. Repeated use of PCR and manipulation of its products cause aerosols that can contaminate neighboring samples and work areas.  Such "carryover contamination" can be prevented by including dUTP in place of dTTP for all amplification reactions.

Detection of Alu by PCR Detection of Alu by PCR - A Human DNA Fingerprinting Lab Protocol - 1994 Cold Spring Harbor Laboratory DNA Learning Center

Differential Display-PCR Protocol Method uses PCR to amplify and display many cDNAs derived from the mRNAs of a given cell or tissue type.  The method relies on two different types of synthetic oligonucleotides: anchored antisense primers and arbitrary sense primers.  A typical anchored primer is complementary to approx. 13 nucleotides of the poly(A) tail of mRNA and the adjacent two nucleotides of the transcribed sequence.

Extraction and Purification of RNA from Tissue-Cultured Cells for Fluorescent mRNA Differential Protocol is the first in a set of three describing fluorescent mRNA differential display (FDD or FDDRT-PCR).  The method begins with the harvesting of total RNA from the tissue-cultured cells of interest.  For other starting materials, such as blood samples, please see Extraction and Purification of RNA from Blood Samples for Fluorescent mRNA Differential Display.

FDD PCR, Identification and Analysis of Differentially Expressed cDNAs Protocol Protocol for FDD PCR, identification and analysis of differentially expressed cDNAs.

Fermentas Taq DNA Polymerase Protocol Fermentas Taq DNA Polymerase Protocol. Fermentas.

General PCR Protocol General PCR Protocol. Sally A. Green

Genetic Engineering with PCR Protocol Method describes how to modify the termini of PCR products by introducing restriction sites and other features.  To reduce the chance of contamination with exogenous DNAs, prepare and use a special set of reagents and solutions for PCR only.  Bake all glassware for 6 hours at 150°C and autoclave all plasticware.

Home-made Taq Polymerase Purification 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

Inverse PCR Inverse PCR. Maddock Lab.

Long and Accurate PCR Protocol Protocol illustrates the rules of successful long PCR: No more than 1 ng of template DNA is used per microliter of PCR in a 100-µl reaction; approximately 0.1 µl of KlentaqLA (not plain Taq) is used per kilobase of target (for targets >10 kb, 1-1.3 µl of enzyme should be used); the Mg++ concentration is considered as the excess over the level of dNTPs.

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

Optimized Welsh DDRT-PCR Protocol Optimized Welsh DDRT-PCR Protocol. Provided by Mirta Grifman and found in Neuroscience Protocols (Nov 1995).

PCR Amplification Manual Roche PCR Amplification Manual Roche. Guide to Standard PCR Protocl, Long PCR Protocol, Hot Start PCR, PCR and Cloning, Basic PCR, other. Roche Applied Science

PCR Cleanup with AMPure magnetic beads PCR Cleanup with AMPure magnetic beads. Charkowski Lab.

PCR Genotyping of Mutant Strains PCR Genotyping of Mutant Strains. Fred Hutchinson Cancer Research Center

PCR Guide - Promega PCR Guide - Promega

PCR Protocol Buffer Creation PCR Protocol Buffer Creation. Based on Jane Gitschier's group.Fred Hutchinson Cancer Research Center.

PCR protocol for cDNA Arrays on Membranes. PDF file. PCR protocol for cDNA Arrays on Membranes. PDF file

PCR Screens of ES Cell Clones Identification of targeted ES cell clones by PCR. Fred Hutchinson Cancer Research Center.

PCR Technology PCR Technology. Informative info on PCR and Protocols. Connie Veilleux