The Student Room Group

Help needed urgently with Gibson Assembly question!!

Hey!

Basically this is the question and I am beyond lost with what’s going on.

‘Design a pair of 40 nt oligonucleotifes to clone the trxA gene into the pET15b expression vector that has been digested with NcoI and XhoI. Restriction digestion is used to prepare the vector. Assembly fragments should overlap by 20 b.p’

I haven’t provided the gene or vector sequence, I just rly need a step by step guide on how to do this. I also don’t understand whether NcoI or XhoI sequences should go at the start or middle of the primer.

Any help would be appreciated ty!

Quick Reply