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DIGITAL PCR-AN EMERGING DISRUPTIVE TECHNOLOGY FOR NUCLEIC ACID REPLICATION
With a higher degree of accuracy and reproducibility in the amplification and analysis of nucleic acids like DNA , RNA and cDNA , this technology is highly promising as compared to the existing forms of PCR
Developed in 1983 , the Nobel Prize winning Polymerase Chain Reaction ( PCR ) has been one of the most commonly used technologies across life science laboratories . Primarily used for amplifying DNA , its application spans cloning , sequencing , analysis of genes , diagnosis and more . Traditional PCR is an end-point analysis . The PCR amplified product is detected by agarose gel electrophoresis making only a semi-quantitative analysis possible . A major drawback with PCR has been that
the quantification of the starting material was never possible . With rapid developments in molecular biology , the need for quantification of DNA in the PCR reaction has been growing .
The technique further improved such that quantitative estimation of DNA became possible and it became today ’ s gold standard for measuring specific amounts of DNA in a sample . Real-time or Quantitative PCR ( qPCR ), is a very powerful and sensitive technique with a broader range of applications compared to PCR , including SNP detection , viral load detection , siRNA / miRNA / RNAi analysis etc .
qPCR utilizes specific complementary oligonucleotide primers that amplify the target of interest along with a fluorescent dye which tracks the progress of the reaction . The reaction requires comparison with a standard curve so as to arrive at the exact concentration of the amplified product . Boehringer Mannheim developed and commercialized first real time PCR . However , qPCR has some limitations such as the need for reference standards and has
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BioVoiceNews | July 2016