PCR - Nagaland Centre
Instrument details:  
Make: Effendorf
Model: 6331 Mastercycler nexus gradient

The PCR involves the primer mediated enzymatic amplification of DNA.

Working principle:The PCR involves the primer mediated enzymatic amplification of DNA. PCR is based on using the ability of DNA polymerase to synthesize new strand of DNA complementary to the offered template strand. Primer is needed because DNA polymerase can add a nucleotide only onto a preexisting 3′-OH group to add the first nucleotide. DNA polymerase then elongate its 3 end by adding more nucleotides to generate an extended region of double stranded DNA. A repetative series of cycles involving template denaturation, primer annealing and extension of the annealed primer by DNA polymerase result in the exponential accumulation of specific fragment whose termini are defined by 5’ end of the primer.

Application:
  • Rapid amplification of tiny fragments of DNA using PCR enabled several techniques such as southern or northern blot hybridization even when the amount of sample material available was very small
  • The major uses of PCR technology include genotyping, gene discovery and cloning, vector construction, transformant, identification, screening and characterization, and seed quality control.
  • For the detection of GM material in a product.
  • Identification of disease susceptible and resistant plants.
  • For study of gene expression patterns is another common application of PCR, where in cells or tissues are analyzed in different stages to check for expression of a specific gene. qPCR can be used here to quantitate the level of gene expression.
  • PCR also assists techniques like DNA sequencing using which segments of DNA from an area of interest can be easily amplified to study genetic mutations and their consequences.
  • The Human Genome Project used PCR to indicate the presence of a specific genome segment in a particular clone. This enabled mapping of the clones and collating results from several laboratories.
  • Advanced variants of the PCR technique have been found to be useful in chromosomal analysis techniques that can help in early detection of genetic birth defects in children.
  • PCR augments the traditional method of DNA cloning by amplifying tiny DNA segments for introduction into a vector. By altering the PCR protocol, site-directed or general mutations can be achieved in the DNA fragment of interest.
User instruction:  
Contact us:

Director, ICAR Research Complex for NEH Region, Umiam 793103, Meghalaya.

Phone No: (0364) 2570257

Email: director.icar-neh@icar.gov.in / icarneh.director@icar.gov.in

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