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Richard Lab:Restriction Digest: Difference between revisions
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Richard Lab:Restriction Digest (view source)
Revision as of 16:14, 10 January 2011
, 10 January 2011→Notes
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*If you're not getting good digestion it might be because your enzyme is bad. Double the digestion time and see. | *If you're not getting good digestion it might be because your enzyme is bad. Double the digestion time and see. | ||
*Longer digest gives more complete digestion, especially if you have >1µg of DNA, but can sometimes give nonspecific digestion | *Longer digest gives more complete digestion, especially if you have >1µg of DNA, but can sometimes give nonspecific digestion | ||
*Beware the the NEB double digest chart. For EcoRI and PstI double digest it recommends using the EcoRI NEBuffer. However based on my digests both NEBuffer 2 and 3 work better; with NEBuffer 2 giving the most complete double digestion. It is funny that | *Beware the the NEB double digest chart. For EcoRI and PstI double digest it recommends using the EcoRI NEBuffer. However based on my digests both NEBuffer 2 and 3 work better; with NEBuffer 2 giving the most complete double digestion. It is funny that they recommend the EcoRI buffer because their chart also says that PstI is only 50% active in that buffer. | ||
==References== | ==References== | ||
