PCR triplicates versus one reaction
Posted 27 March 2011 - 05:08 PM
Posted 28 March 2011 - 09:33 PM
Run your PCR in gradient.
You have to do all the above (unless I missed out something...) in order to answer your question. You will know the minimum template DNA required for that particular PCR to work. No general or common or fixed number of amount.
I not sure what you mean by PCR bias. Bad primer design: unspecific amplification.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
Posted 31 March 2011 - 07:36 AM
i don't understand how you can dilute the dna without decreasing the starting template amount. can you give some examples.
If you put 50ng in 50ul, you have a concentration of 1ng/ul. If you put 50ng in 100ul, you have a concentration of 0.5ng/ul but you still have 50ng in your tube.
As for the low copy number PCR, we usually increase the number of cycles in order to detect PCR product on a gel.