I have done an extraction for DNA from amniotic fluid sample using Thermo Kit but when i measured DNA on the Nano drop I realized that it is contaminated sample as the ratio 260/280 is 0.7..I tried to make PCR with concentration 100ng in total volume 15 microlitre but no bands appeared. could you please tell me how can I increase the purity of sample and how to overcome this problem to get a successful PCR?
Thanks
Does the kit use phenol? If yes, any carry-over will decrease the 260/280 ratio.
A high amount of proteins in your sample can also cause a low ratio.
100ng of DNA in only 15 microliters is a LOT of DNA. I know it sounds counter-intuitive, but try diluting your DNA (1:10, 1:100, 1:1000). Try all 3 dilutions + controls.
Dilution can lower the amount of PCR-inhibiting proteins, but the polymerase is sensitive enough to still amplify the DNA.
Good luck!
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