Top : Molecular Biology : PCR

Summary: Polymerase Chain Reaction (PCR), invented by Kary B. Mullis, at the Cetus Corporation, who was awarded the 1993 Nobel Prize for chemistry for PCR, is a technique to exponentially amplify in vitro a small quantity of a specific nucleotide sequence in the presence of template sequence, two oligonucleotide primers that hybridize to opposite strands and flank the region of interest in the target DNA, a thermostable (taq) DNA polymerase. The reaction is cycled involving template denaturation, primer annealing, and the extension of the annealed primers by DNA polymerase until enough copies are made for further analysis.
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Protocols
  • Complete PCR Guide (Laboratory of Dr. Bart Frank, Oklahoma Medical Research Foundation)
    In the polymerase chain reaction (PCR), a thermostable DNA polymerase amplifies DNA that is flanked by known sequences. The known sequences correspond to those on synthetic oligonucleotide primers which are used to initiate the reaction. PCR can be used in many complex ways to achieve different results.
    http://www.omrf.org/omrf/research/09/FrankLab/PCR....
    Added: Sat Jan 31 2009, Hits: 14918, Reviews: 0  Write review   Cached
  • Home-Made Taq Polymerase Purification (Dr. Simon Dawson, Department of Biochemistry, University of Nottingham Medical School)
    Taq polymerase can be expensive. Here is the protocol for making your own Tag polymerase and it gives stable results.
    http://www.nottingham.ac.uk/~mbzspd/methods/Home_m...
    Added: Sat Apr 10 2004, Hits: 435, Reviews: 0  Write review   Cached
  • Interactive guide for solving common PCR-problems (Roche Diagnostics Corporation)
    Troubleshooting hints based on the five most common symptoms observed: No product / Misincorporation or low fidelity / Non-specific bands / Smeared bands/Low yield.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 1064, Reviews: 0  Write review   Cached
  • Quantitative RT-PCR and Other PCR Procedures (Jack Vanden Heuvel, Penn State University)
    A must-read protocol for PCR beginners as well as verterans. It touches in depth aspects specific to quantitative RT-PCR as well as those common to standard PCR.
    http://www.cas.psu.edu/docs/CASDEPT/VET/jackvh/jvh...
    Added: Sun Mar 28 2004, Hits: 8878, Reviews: 0  Write review  
  • Calculating Concentrations for PCR (Molecular Biology Techniques Manual)
    How to calculate primer and nucleotide concentration
    http://web.uct.ac.za/microbiology/pcrconcn.htm
    Added: Tue May 14 2002, Hits: 5713, Reviews: 0  Write review   Cached
  • Guidelines for determining the number of PCR cycles (Qiagen)
    http://www.qiagen.com/resources/info/guidelines_fo...
    Added: Sun Jul 21 2002, Hits: 3440, Reviews: 0  Write review   Cached
  • PCR and Agarose Gel Protocol (Genomics Facility, University of Edinburgh)
    Describes PCR reaction and running agarose gel
    http://staff.biology.ed.ac.uk/services/equipment/g...
    Added: Sat Jan 31 2009, Hits: 5107, Reviews: 0  Write review   Cached
  • PCR clean up using 3M sodium acetate and chilled absolute ethanol (KAIMRC, NGHA )
    Clean up PCR in 1.5 hrs. with a very easy steps.
    Added: Sat Jan 02 2010, Reviews: 0  Write review  
  • PCR reaction components (Roche Diagnostics Corporation)
    Very useful introduction to PCR componets including template, primers, dNTP, additive, polymerase, pH, and MgCl2 concentration.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 770, Reviews: 0  Write review   Cached
  • Preventing carry-over contamination with uracil-DNA glycosylase (Roche Diagnostics Corporation)
    PCR can amplify a single molecule over a billionfold. Thus, even minuscule amounts of a contaminant can be amplified and lead to a false positive result. Such contaminants are often products from previous PCR amplifications (carry-over contamination). One common strategy to avoid such contamination is substituting dUTP for dTTP during PCR amplification, to produce uracil-containing DNA (U-DNA).
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 213, Reviews: 0  Write review   Cached
  • Primer Extension PDF file (Cross Lab, Rockefeller University)
    Protocol that uses Promega Primer Extension system
    http://tryps.rockefeller.edu/Protocols/primer_exte...
    Added: Sat Jan 31 2009, Hits: 2663, Reviews: 0  Write review  
  • Setting up the laboratory to avoid contamination (Roche Diagnostics Corporation)
    The ability of the Polymerase Chain Reaction to amplify a single molecule means that trace amounts of DNA contaminants could serve as templates, resulting in amplification of the wrong template (false positives). This guide describes the common sources of contamination such as lab facilities and sample handling, and how to avoid contamination.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 432, Reviews: 0  Write review   Cached
  • Thermal Cycling Profile for Standard PCR (Roche Diagnostics Corporation)
    Basic guide to PCR cycling and how to optimize each step.
    http://www.roche-applied-science.com/pcr/applicati...
    Added: Sat Sep 21 2002, Hits: 591, Reviews: 0  Write review   Cached
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