Caister Academic Press

Polymerase Chain Reaction PCR Protocols

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A collection of PCR protocols that are available free online. The protocols are taken from published papers in peer-reviewed journals or from edited books. They are provided here as a resource to aid scientists that are using or developing PCR techniques in the laboratory.
  1. Core-Sampling a PCR Band and Reamplification
    The core sample is a defined volume, may be stored indefinitely, and provides material for multiple re-amplifications.
  2. One-step DNA Fragment Assembly and Circularization for Gene Cloning
    A one-step procedure based on Taq polymerase for the precise assembly of DNA fragments into circular constructs as long as 6 kb.
  3. SPUD qPCR Assay Confirms PREXCEL-Q Software's Ability to Avoid qPCR Inhibition
    Avoid qPCR inhibition using PREXCEL-Q sample and standard dilution calculations.
  4. Efficient Cloning of Alternatively Polyadenylated Transcripts via Hybridization Capture PCR
    A simple and rapid approach combining SMART technology for the construction of a full length cDNA library and hybrid capture PCR for the selection and amplification of target cDNAs. The strategy is characterized by enhanced specificity compared to other conventional RT-PCR and 3' RACE procedures.
  5. Endonuclease-Mediated Long PCR and Its Application to Restriction Mapping
    Restriction digestion performed prior to long PCR amplification can be used to selectively suppress the amplification of members of families of closely related DNA sequences, thereby making it possible to selectively amplify one of a group of highly homologous sequences.
  6. Molecular Diagnosis of Medical Viruses
    This review discusses the applications of qualitative and quantitative real-time PCR, nested PCR, multiplex PCR, nucleotide sequence analysis of amplified products and quality assurance with nucleic acid testing (NAT) in diagnostic laboratories.
  7. A PCR-based Method for Isolation of Genomic DNA Flanking a Known DNA Sequence
    A simple PCR-based method for the isolation of genomic DNA that lies adjacent to a known DNA sequence.
  8. PCR Clamping
    An efficient, PCR based method for the selective amplification of DNA target sequences that differs by a single base pair. The method utilises the high affinity and specificity of PNA for their complementary nucleic acids and that PNA cannot function as primers for DNA polymerases.
  9. Universal TA Cloning
    The same T-vector can be used to clone any double-stranded DNA fragment, including PCR products. This is especially useful when compatible restriction sites are not available for the subcloning of DNA fragments from one vector to another. Directional cloning is made possible by appropriate hemi-phosphorylation of both the T-vectors and the inserts. Any DNA fragment can be cloned without compromising the cloning efficiency making the universal TA cloning method convenient and labour-saving.
  10. Analysis of Specific Bacteria from Environmental Samples using a Quantitative Polymerase Chain Reaction
    The use of quantitative PCR for measuring bacterial abundance in environmental samples. Two different approaches are described.
  11. DNA Splicing by Directed Ligation (SDL)
    Splicing by directed ligation (SDL) is a method of in-phase joining of PCR-generated DNA fragments that is based on a pre-designed combination of class IIS restriction endonuclease recognition and cleavage sites.
  12. Gene Amplification from Cryopreserved Arabidopsis thaliana Shoot Tips
    This protocol will help researchers to pursue faurther research in the field of molecular genetics of plant cryostress. Interesting genes identified via these process can be cloned and plants can be transfomred for the purpose of trait enhancement and modification.

Further reading