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Simplified detection of the asymmetric polymerase chain reaction-amplified DNA and its application in the target identification

G Suhasa, Savithri Bhat.




Abstract
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The nucleic acid amplification methods such as the PCR and the LAMP rely on the detection of the amplified products by means of agarose gel electrophoresis. The detection of the amplified target DNA in an asymmetric PCR was simplified by carrying out the probe hybridization, polymerization and the subsequent measurement of fluorescence of the dsDNA by using the Qubit® dsDNA BR assay kit. This method was validated by detecting the lacZ gene that is present in the pUC 18 plasmid as well as by detecting the CDH13 gene that is present in the human genomic DNA.

Key words: Polymerase chain reaction, agarose gel electrophoresis, asymmetric PCR, loop mediated isothermal amplification, fluorometry.






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