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  • Standard PCR Protocol (Molecular Biology Techniques Manual)
    The followings are described in detail
    Recommended Reagent Concentrations
    Recommended Reaction Conditions
    "Hot Start" PCR
    Asymmetric PCR for ssDNA Production
    Detecting Products
    Labeling PCR Products with Digoxigenin
    Cleaning PCR Products
    Sequencing PCR Products
    Cloning PCR Products
    http://web.uct.ac.za/microbiology/pcrcond.htm#Labe...
    Added: Tue May 14 2002, Hits: 4326, Reviews: 0  Write review   Cached
  • Standard PCR Protocol (Molecular Biology Techniques Manual)
    The followings are described in detail
    Recommended Reagent Concentrations
    Recommended Reaction Conditions
    "Hot Start" PCR
    Asymmetric PCR for ssDNA Production
    Detecting Products
    Labeling PCR Products with Digoxigenin
    Cleaning PCR Products
    Sequencing PCR Products
    Cloning PCR Products
    http://web.uct.ac.za/microbiology/pcrcond.htm#Labe...
    Added: Tue May 14 2002, Hits: 12636, Reviews: 1 Read reviews  Write review   Cached
  • Degenerate PCR (Atle M. Bones Lab, Norwegian University of Science and Technology)
    Degenerate PCR is in most respects identical to ordinary PCR, but with one major difference. Instead of using specific PCR primers with a given sequence, you use mixed PCR primers. That is, if you do not know exactly the sequence of the gene you are going to amplify, you insert "wobbles" in the PCR primers where there is more than one possibility. For instance, if you just have a protein motif, you can back-translate the protein motif to the corresponding nucleotide motif. (Protein --> Sequence).
    http://boneslab.bio.ntnu.no/degpcrshortguide.htm
    Added: Sun Mar 22 2009, Hits: 8683, Reviews: 0  Write review   Cached
  • PCR and multiplex PCR guide (Octavian Henegariu, Yale University)
    Detailed guide for PCR optimization, like concentration of various components, cycling conditions, adjuvants and more.
    http://info.med.yale.edu/genetics/ward/tavi/Guide....
    Added: Tue May 14 2002, Hits: 19666, Reviews: 0  Write review   Cached
  • An Economic PCR Enhancer for GC-Rich PCR Templates (Protocol Online)
    An combinatorial enhancer solution (CES) which efficiently improves many PCR using different polymerases.
    Added: Mon Feb 02 2009, Reviews: 0  Write review  
  • Confirmation PCR (Saccharomyces Genome Deletion Project)
    To study the genetics of yeast, yeast gene ORF is deleted by PCR based strategy to generate mutated strain. Confirmation PCR is used to verify if the yeast ORF deletion is successful Introduction/General strategy
    Confirmation PCR primer design
    Primer organization in the 96-well plates
    Oligonucleotide concentrations
    Quality control procedures
    Single tube PCR protocol
    96-well format protocol
    http://sequence-www.stanford.edu/group/yeast_delet...
    Added: Tue May 14 2002, Hits: 1865, Reviews: 0  Write review   Cached
  • Methylation-specific PCR: a novel PCR assay for methylation status of CpG islands (PNAS)
    This original article by James Herman and Stephen B. Baylin et al. first described the MSP method which has since become popular for DNA methylation screening.
    http://www.pnas.org/content/93/18/9821.abstract
    Added: Tue Feb 03 2009, Hits: 4386, Reviews: 0  Write review   Cached
  • PCR Program Design (Octavian Henegariu)
    For any given primer pair, the PCR program can be selected based on the composition (GC content) of the primers and the length of the expected PCR product. Here is a guide to select the cycling program of PCR.
    http://info.med.yale.edu/genetics/ward/tavi/p08.ht...
    Added: Tue May 14 2002, Hits: 4708, Reviews: 0  Write review   Cached
  • Purification and Sequencing of PCR Product (Donis-Keller lab)
    This method is used to sequence directly a specific PCR product after large scale amplification. The reaction should be carried out in the usual fashion, except that after optimization of annealing temperature and other conditions in the 5 µl volume, a large scale 50 µl (10X) total reaction volume should be used to generate sufficient PCR product for sequencing.
    http://hg.wustl.edu/hdk_lab_manual/pcr/pcr3.html
    Added: Tue May 14 2002, Hits: 2805, Reviews: 0  Write review   Cached
  • Purification and Sequencing of PCR Product (Donis-Keller lab)
    This method is used to sequence directly a specific PCR product after large scale amplification. The reaction should be carried out in the usual fashion, except that after optimization of annealing temperature and other conditions in the 5 µl volume, a large scale 50 µl (10X) total reaction volume should be used to generate sufficient PCR product for sequencing.
    http://hg.wustl.edu/hdk_lab_manual/pcr/pcr3.html
    Added: Tue May 14 2002, Hits: 1294, Reviews: 0  Write review   Cached
  • Cleavage Efficiency Close to the Termini of PCR Fragments (Fermentas)
    When designing PCR experiments in which the synthesized DNA fragment is to be subsequently digested with a RE, it is very important to determine how many extra nucleotides should be added to the 5’-end of the PCR primer next to the introduced recognition site to ensure efficient cleavage with the appropriate restriction endonuclease. Some restriction endonucleases cleave DNA poorly when their recognition sites are located at the ends of DNA fragments. Information on restriction endonuclease performance close to the end of DNA fragments may be helpful when planning the introduction of cleavage sites at DNA termini in PCR experiments.
    http://www.fermentas.com/techinfo/RE/restrdigpcrii...
    Added: Tue May 14 2002, Hits: 382, Reviews: 0  Write review   Cached
  • Cleavage Efficiency Close to the Termini of PCR Fragments (Fermentas)
    When designing PCR experiments in which the synthesized DNA fragment is to be subsequently digested with a RE, it is very important to determine how many extra nucleotides should be added to the 5’-end of the PCR primer next to the introduced recognition site to ensure efficient cleavage with the appropriate restriction endonuclease. Some restriction endonucleases cleave DNA poorly when their recognition sites are located at the ends of DNA fragments. Information on restriction endonuclease performance close to the end of DNA fragments may be helpful when planning the introduction of cleavage sites at DNA termini in PCR experiments.
    http://www.fermentas.com/techinfo/RE/restrdigpcrii...
    Added: Tue May 14 2002, Hits: 419, Reviews: 0  Write review   Cached
  • Troubleshooting for PCR and Multiplex PCR (Octavian Henegariu, Yale University)
    http://info.med.yale.edu/genetics/ward/tavi/Trbles...
    Added: Thu Apr 22 2004, Hits: 9156, Reviews: 0  Write review   Cached
  • PCR (Chen)
    Standard PCR protocol with detailed notes on every aspects of PCR, such as cycling conditions, components used, additives, and primer design.
    http://www.biochem.ucl.ac.uk/~chen/protocols/PCR.h...
    Added: Tue May 14 2002, Hits: 21332, Reviews: 0  Write review  
  • PCR Mutagenesis (Matt Lewis, Department of Pathology, University of Liverpool)
    This method uses a proof-reading polymerase to read all the way around a plasmid and thus incorporate the primer as the new (mutant) sequence. Only a few (say 12) PCR cycles are performed on a relatively large amount of plasmid template to minimise the chance of expanding PCR sequence errors.
    http://www.methodbook.net/pcr/pcrmut.html
    Added: Fri Sep 06 2002, Hits: 6837, Reviews: 0  Write review   Cached
  • Inverse PCR PDF file (Langdale Lab, Department of Plant Sciences, University of Oxford)
    Another two methods from the same laboratory are also available at http://dps.plants.ox.ac.uk/langdalelab/protocols/PCR/iPCR_Susie.pdf and http://dps.plants.ox.ac.uk/langdalelab/protocols/PCR/iPCR_Tom.pdf
    http://dps.plants.ox.ac.uk/langdalelab/protocols/P...
    Added: Wed Feb 11 2009, Hits: 6956, Reviews: 0  Write review  
  • Purification and Sequencing of PCR Product (Donis-Keller lab)
    This method is used to sequence directly a specific PCR product after large scale amplification. The reaction should be carried out in the usual fashion, except that after optimization of annealing temperature and other conditions in the 5 µl volume, a large scale 50 µl (10X) total reaction volume should be used to generate sufficient PCR product for sequencing.
    http://hg.wustl.edu/hdk_lab_manual/pcr/pcr3.html
    Added: Tue May 14 2002, Hits: 1354, Reviews: 0  Write review   Cached
  • What's PCR? (Michael Blaber's Lab)
    Illustrated introduction to various aspects of PCR technique. It's very valuable not only for those new to PCR, but also those familiar with PCR.
    http://wine1.sb.fsu.edu/bch5425/lect22/lect22.htm
    Added: Tue May 14 2002, Hits: 691, Reviews: 0  Write review   Cached
  • Site-directed Mutagenesis using PCR (NWFSC)
    Use of PCR in site-directed mutagenesis accomplishes strand separation by using a denaturing step to separate the complementing strands and allowing efficient polymerization of the PCR primers. PCR site-directed methods thus allow site-specific mutations to be incorporated in virtually any double-stranded plasmid; eliminating the need for M13-based vectors or single-stranded rescue.
    http://micro.nwfsc.noaa.gov/protocols/pcr-mut.html
    Added: Tue May 14 2002, Hits: 2163, Reviews: 0  Write review  
  • Genome walking by PCR PDF file (Langdale Lab, Department of Plant Sciences, University of Oxford)
    The protocol involves a nested PCR with a touch down program. This method is very prone to artefacts so you NEED to follow rigorously the requirements for the design of primers and the touch-down PCR program!
    http://dps.plants.ox.ac.uk/langdalelab/protocols/P...
    Added: Wed Feb 11 2009, Hits: 1868, Reviews: 0  Write review  
  • 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: 15048, Reviews: 0  Write review   Cached
  • PCR from Spores (Fungal Genetics Stock Center)
    a PCR protocol which uses M. grisea conidia directly for PCR analysis without extraction of DNA.
    http://www.fgsc.net/fgn42/xu.html
    Added: Mon Dec 09 2002, Hits: 1520, Reviews: 0  Write review   Cached
  • Real-Time or Kinetic PCR (University of Iowa DNA Facility)
    This page describes the following topics on real-time PCR:
    • Real-Time Chemistry
    • Instrumentation
    • Real-Time PCR Quantitation
    • Primer and Probe Design 
    • Thermal Cycling Parameters 
    • Sample Preparation
    • Data and Analysis
    • Supplemental Information

    http://dna-9.int-med.uiowa.edu/realtime.htm
    Added: Sun Sep 22 2002, Hits: 7273, Reviews: 0  Write review   Cached
  • PCR (Julie B. Wolf, University of Maryland, Baltimore County)
    PCR components and cycling conditions
    http://www.research.umbc.edu/~jwolf/m2.htm#pcr
    Added: Tue May 14 2002, Hits: 12068, Reviews: 0  Write review   Cached
  • RT In Situ PCR (Gerard J. Nuovo)
    RT in situ PCR allows for the routine and rapid detection of low copy viral and human RNAs. Success with RT in situ PCR is best accomplished with formalin fixed, paraffin embedded material, which allows the study of archival material.
    http://www.bioscience.org/1996/v1/c/nuovo1/htmls/l...
    Added: Tue May 14 2002, Hits: 2404, Reviews: 0  Write review   Cached

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