pUC19 ligation - (Oct/20/2008 )
I have a quick question about ligating random segments into pUC19. I have a protocol and it says cut the DNA in question with SauA1. Then cut the vector with BamH1. I thought you had to cut the DNA and vector with the same enzyme so that the two sticky-ends match up?
-shayn-
Hi Shayn,
I presume you are using Sau3AI. The restriction nzymes BamHI and Sau3Al sites have identical cohesive ends. So The vector digested with BamHI and Insert digested with BamHI have compatible cohesive ends, so that these two DNA can be ligated.
Bam HI site 5' G/GATCC 3'
Sau3A1 5' /GATC 3'
/ indicates the cut site
Regards
aviprem
http://aviprem.gq.nu
QUOTE (shayn @ Oct 21 2008, 11:15 AM)
I have a quick question about ligating random segments into pUC19. I have a protocol and it says cut the DNA in question with SauA1. Then cut the vector with BamH1. I thought you had to cut the DNA and vector with the same enzyme so that the two sticky-ends match up?
-aviprem-