Restricition enzyme qouted in paper doesnot cut the PCR product according to the - (Nov/28/2007 )
hello everyone.
This is the sequence where my PCR product is supposed to be cut by restricition enzyme: ....ACGC... One paper i consulted mentions that enzyme HpyCH4IV cuts it at this point. but the restriction site this enzyme recognizes is 5`...ACGT...3` {and 3`...TGCA...5` }. Thus understandably when i enter the sequence for my PCR product in "NEBcutter" it gives a list of enzymes which can cut it. HPYCH4IV is not one of them. NEBcuuter actually mentions a different enzyme HpyAV which cuts at this point.
What is going on? Could HPYCH4IV cut my PCR product even when one out of four bases doesnot match? Could the paper i read have made a typing mistake . seems unlikely as it is from a reputed europian journal.
giudance please
I would say there is a typo error in the paper.
HpyCH4IV most certainly cuts at 5`ACGT 3`
Or an error in the sequence.
hello everyone,
just thought i'd inform everybody that i found the solution to my problem. The primers quoted in the paper are mutation primers which produce a restricition site in the PCR product not present in the original gene itself.