why concentrated DNA cannot be used in PCR

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ANS: DNA consist of Adenine, Thymine, cytosine and Guanine .There are two hydrogen bonds between A and T ,three hydrogen bonds between G and C ,the presence of more hydrogen bonds between G and C , the amplification of GC rich DNA is not occur in proper way ,if the concentration of GC content increases the melting temperature of the reaction and difficult to amplify. About 30ng to 50 ng DNA is required for 25 microliter PCR reaction.suppose the concentration of DNA is low then DNA might not be amplified and higher concentration of DNA hinder s the PCR reaction .Two conditions may occur DNA may amplify or DNA may not amplified.

The higher concentration of DNA ,the PCR tube filled with higher amount of nonspecific DNA and hinders in the activity of other reagents and may not reach at the target DNA sequence that may lead to non amplification of DNA.

In other possibility, due to the higher amount of non-target template DNA ,the primer bind to other than its complementary sequences and result in wrong amplification.

gladiator answered 1 year ago

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