I want to use a short stretch of single stranded DNA, say, from 40nts to 75nts. I want to amplify this sequence using PCR. Is it possible to amplify such short sequences using PCR.
for 40mer it will better to purchase it like u do PCR primers ..
If you want to amplify ssDNA you can do assymetric PCR ..In doing your PCR use one primer in large excess
Dear joy,
Can u please tell what further downstream process, you want to perform with your PCR product....
For 60 and higher you could easily use PCR for amplification. You will have to play with conditions and reagents though to find the optimal ones. In addition, PCR reaction may produce some bi-products (non-specific DNA)I that may be removed by purification via PAAG electrophoresis followed by elution/extraction and EtOH precipitation. For production of shorter sequences, I would use a synthetic ssDNA template and either extend it by primer extension method (with Klenov) after annealing to the short (18-20 nt) primer or simply hybridize it to the complementary strand. In both cases, the final dsDNA should be gel-purified (PAAG not agarose).
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