what is your DNA concentration after gel extraction?
Posted 11 June 2012 - 12:48 AM
Is it always like this? I keep losing a lot of DNA, PCR product or digested plasmids. There are also other kits which are not column based, like Qiagen's Qiaex II, but you lose a lot of DNA with them too.
what is your concentration after gel extraction?
Posted 11 June 2012 - 04:41 AM
Posted 11 June 2012 - 05:55 PM
Phage, may I know what method you use if you want to avoid gel extraction?
Bob1, I also had no problem with having 10 ng/ul previously, because as you said we don't need much DNA for cloning and ligation. But now I am struggling with Overlap Extension PCR and I need high amount of template DNA for the second PCR. I cannot add more than 10-15 ul of the purified PCR product to my second PCR, and I am worried if this is the reason my PCR fails.
Posted 11 June 2012 - 06:12 PM
Assembly of BioBrick standard biological parts using three antibiotic assembly.
Shetty R, Lizarazo M, Rettberg R, Knight TF.
Methods Enzymol. 2011;498:311-26. PMID: 21601683