@VafaOmid @TheMenacheryLab There are probably unidentified batCovs with s1/s2 furin sites. That's my point. Also, with good recombination machinery you can get away very small overlaps. All yeast need are 20bp on either flank. https://t.co/p8F687meZS
The only reason Gibson assembly is a thing is because Daniel Gibson realized an inferior version of yeast recombination using enzymes is patentable. Seriously. Anything Gibson can do, yeast can do a million times better. https://t.co/27RFTMTMsF
@DanutaSastre @Caroline_Bartma You use a yeast vector with a selectable marker (usually covering auxotrophy- no worrying about satellite colonies). PCR out fragments with 30bp of homology to each adjacent segment, transform into yeast with LiAc method, …
#Bio140 #UCMerced Gap-repair cloning is a highly efficient way to fix errors in DNA https://t.co/UiIOo1CRVO - Antonio S.