How can I double digest pET 28a vector with BamHI and HindIII enzymes as their sites in vector are very close to each other?

05 May 2018 1 2K Report

I have tried to digest pET 28a vector with BamHI and HindIII enzymes but didn't get any band rather very light multiple bands were appeared in the gel. How can I be able to double digest my vector as I am only restricted to use these enzymes.

Mohammed Al-Nussairawi

YOU CAN

use the following protocol

Tango buffer 2 microL

MQ water 7.7 microL

DNA template 10 micro L

HINDIII 0.3-0.5 microL

BamHI 0.3-0.5 microL

AT 37 C for 2h and try at 37C for 40 minutes as well

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