Category Matches: 27
Protocol Matches: 282
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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: 4415, Reviews: 0
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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: 12728, Reviews: 1 Read reviews
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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: 8877, Reviews: 0
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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: 19810, Reviews: 0
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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
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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: 1924, Reviews: 0
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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: 4498, Reviews: 0
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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: 4783, Reviews: 0
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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: 2892, Reviews: 0
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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: 1367, Reviews: 0
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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: 383, Reviews: 0
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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: 420, Reviews: 0
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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: 9271, Reviews: 0
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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: 21431, Reviews: 0
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- 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: 6908, Reviews: 0
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Cached -
Inverse PCR
(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: 7045, Reviews: 0
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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: 1425, Reviews: 0
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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
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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: 2239, Reviews: 0
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Genome walking by PCR
(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: 1937, Reviews: 0
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- 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: 15222, Reviews: 0
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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: 1600, Reviews: 0
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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
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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: 12151, Reviews: 0
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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: 2463, Reviews: 0
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