restriction enzyme problem - restriction enzyme problem (Jun/09/2009 )

hi admin and chatter,

today, i'm using running re to cut my sequence, yes the re was cutting my sequence, my problem is the re was cutting at wrong site. base on http://tools.neb.com/NEBcutter2/index.php, b4 i order the RE, the re should cutting 123, 225 (example). but now base on my gel picture ..the RE not cutting what we aspect in NEB.. i'm using 100 bp marker.

pls give some opinion if u had same problem.. some solutions... thank you

you may be seeing star activity.

can we see the gel image please? And what enzyme you're using.

more detail, actually i'm doing RE digestion of PCR products.

you can look at a similar (same?) problem's responses in the biotechniques forum.

pay particular attention to the response made by onyourcase.