ligatin issues - (Jul/28/2009 )
Hi all,
I want to clone 3kb gene into PTZ57R/T vector.but it is not getting ligated. after ligation i could see only the insert and not the ligated product.
what is the maximum size of gene that can be cloned into PTZ57R/T vector.
seggestions needed
thanx
-novagen-
novagen on Jul 28 2009, 11:48 AM said:
Hi all,
I want to clone 3kb gene into PTZ57R/T vector.but it is not getting ligated. after ligation i could see only the insert and not the ligated product.
what is the maximum size of gene that can be cloned into PTZ57R/T vector.
seggestions needed
thanx
I want to clone 3kb gene into PTZ57R/T vector.but it is not getting ligated. after ligation i could see only the insert and not the ligated product.
what is the maximum size of gene that can be cloned into PTZ57R/T vector.
seggestions needed
thanx
Never used PTZ57R/T vector so not sure about max sizes (check manual/supplier). As for checking ligation its really not worth checking ligation on a gel, the concentration of insert/vector can be very low to see on a gel and no matter how hard to try you'll never get 100% ligation efficiency anyway. Its better to just run controls with your cloning (vector no insert) and see how many colonies if you get loads then its religated vector in which case punt for some shrimp alkaline phosphatase.
hope that helps somewhat.
-uncutlateralus-