not getting transformants - (Aug/20/2007 )
You meant that when you designed primers, there were no add-on bases in front of the NdeI site, for example:
5'-AAAAAACATATGNNNNNNN (the N sequence is your gene sequence) -3'
If there is no add-on, the efficiency of digestion of your PCR product is greatly reduced. That counts for both enzymes, not only NdeI. But of course some enzymes are pickier than others.
-Almasy-
for NdeI is even worst.
5'-CATATGNNNNNNN (the N sequence is your gene sequence) -3'
Without any skirting, the NdeI site will not cut. NdeI is one of the picker enzymes.
-perneseblue-
QUOTE (perneseblue @ Sep 10 2007, 04:32 AM)
for NdeI is even worst.
5'-CATATGNNNNNNN (the N sequence is your gene sequence) -3'
Without any skirting, the NdeI site will not cut. NdeI is one of the picker enzymes.
5'-CATATGNNNNNNN (the N sequence is your gene sequence) -3'
Without any skirting, the NdeI site will not cut. NdeI is one of the picker enzymes.
thank you
i will go for new primers as you said and iwill try that
before going to order any other precautions to fallow in addition for better result
please suggest
thank you
-janu-