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| kramer |
| Jenn's email question about subcloning |
| 2009.04.06 09:00:20 | |
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This web site is seriously weak...doesn't work with IE7...only Mozilla. Here is a cut-and-paste of an email conversation with Jenn about the subcloning exercise for BIO212AA S'09.
body {margin:8px} .tr-field {font:normal x-small arial} Jen, Sorry so late in replying. Daniel was walking through the same logic today with me, so I'll tell you the conclusions we came to... The pUC19 MCS is in the LacZ operon, so any changes there will make the gene inoperative. The cut is 23 nucleotides long so one would hope that if you stuck 23 "new nucleotides" in...well, at least there wouldn't be a frame shift mutation. I don't know that it's reasonable to expect proper expression of LacZ though...so it's anyone's guess whether blue-white selection will work any more. (I think not.) Re: polymerase synthesizing the genes...no, I don't think it understands content, but if you have nonsense nucleotide sequences, the synthesized protein won't work (reference blue-white selection discussion above). Soooo...yeh. I think you're right. If you successfully clone something else in there, LacZ won't work anymore. I'm going to cut-and-paste this discussion into the bio-blog-o-sphere (has the language really denegrated to THAT?) Peace, _ C On Fri, Apr 3, 2009 at 1:06 PM, jennifer kistner <
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> wrote:
Tags: BIO212AA subcloning pUC19 LacZ
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