I am trying to digest my plasmid(extracted by kit) using SfiI and NotI enzymes but even changing many conditions, it goes not work at all....some of thimgs which i have tried so dar are given below.
Double digestion using fast digest enzyme and buffer.
Overnight digestion using normal enzymes ( not fastdigest).
I even tested my enzymes on other samples but it works fine with them.
Tried to digest the PCR products after amplification but no colony after transformation.
Any suggestion will be appreciated.
Note...i used 1 microgram of plasmid
Restriction digestion with NotI and SfiI