While amplifying particular part of a gene, we are adding genomic DNA. But I observed only PCR products not genomic DNA. Why genomic DNA is not appeared in PCR product?
Normally, the amount of amplified DNA is so vastly larger than the template DNA originally present in a PCR reaction that the latter is not detectable on a gel if the reaction went OK and if the gel is not grossly overloaded
most of the genomic DNAs are in the well, they are too big to run. if the concentration of the genomic DNA is low, they are difficult to notice in the well.
What you are seeing is the normal and expected result. You would not expect to see any gDNA on an agarose gel after PCR to amplify just one region. Too few copies of the gDNA are present to be visible.
May be to different reasons such as:
•Contamination of reagents or lab, results in false positive results.
•Failure due to a mistake in the protocol.
•Different materials/parts of the sample can inhibit the PRC process.
• Inhibitors,’ which interfere or prevent the DNA amplification process from occurring properly.
•These inhibitors can be present in DNA samples.
•Inhibitors can:
(1)interfere with the cell lysis necessary for DNA extraction,
(2) interfere by nucleic acid degradation or capture,
inhibit polymerase activity,
As others have said, there is a low concentration of genomic DNAs in the well, which is big to run in agarose gel.
Genomic DNA will denaturated to form two seperated strand and as its very long Chain its imposible to recombine, the Dye will conjugated with bond between double strand only
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