HELP! Posted November 1, 2008 Share Posted November 1, 2008 I have done an experiment with restriction enzymes, single and double and triple digestions. I have the size of all the fragments in base pairs, how do I locate the restriction sites? many many many thanks to anyone who can help!! Link to comment Share on other sites More sharing options...
ecoli Posted November 1, 2008 Share Posted November 1, 2008 I use NebCutter: http://tools.neb.com/NEBcutter2/index.php You have the sequences, right? Link to comment Share on other sites More sharing options...
HELP! Posted November 3, 2008 Author Share Posted November 3, 2008 NO!! I think this is where the problem lies, I only have the sizes of the fragements (bp) generated by the digests! Link to comment Share on other sites More sharing options...
CharonY Posted November 3, 2008 Share Posted November 3, 2008 Well, without the sequence you won't know what fragment sizes to expect. Assuming perfect restriction and depending on the combination of restriction you can get some information regarding the DNA you cut, though. The basic question is whether you got linear or circular DNA, then take a digest to map out the overall size and then map the fragments from each digest back to the original DNA fragment (as the sum of all fragments is always the molecule size). Link to comment Share on other sites More sharing options...
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