I am doing restriction digestion for TSP45i and ScrfI enzymes with PCR product of plant genomic DNA . Not able to, differentiate between before and after restriction digestion..
Partial diegtion menas....If you are getting a 400 bp band ...you should also see a 300 bp band and an uncut 700 bp band....if you don't mind can you send a photograph of the results you mentione above..
If you suspect partial digestion you can do following things...
1) purify the PCR product as some enzymes are sensitive to impurities
2) check the buffer and temp conditions of the enzymes
3) increase the units of enzyme or else the incubation time...bye
Thank you ...sure i will send you gel photo....it's 150 bps uncut last 4- wells and ...starting wells with two bands of 150 and 130.....let me know the reason.....