I have isolated gDNA from human dried spots with a yield of 13.8ng/uL and a purity (260/280) of 1.82. However I observe that these isolated DNA is not amplifying in PCR. Can anyone tell me why this is? What can I do to ensure amplification of my isolated DNA?
What protocol are you using for amplification? Different protocols are specific for amount of DNA you need to load and for the efficiency of the enzyme.
You did not mention how many ul you used for amplification. normally 10 to 50ng should be enough. Did any of your positive control worked? i.e., other PCR procedure is correct, also negative control?
Hey Subhash, Thanks for the input, I used 4uL (~40ng)of the DNA template for amplification. Yes, my NTC and Pos. control worked.
Hey Elias,
I am amplifying the DNA templates to do a downstream sequencing. The expected product size is 2219bp.
Hey Yermia,
I used two protocols. One based on a salting out method and the other using a QIAamp DNA mini kit.
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