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how to amplify Red/ET and Cre expression plasmid

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wildaisy
United States

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Topic Started by wildaisy
on 5/1/2009 7:30 AM
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I am working with a BAC kit and want to amplify the Red/ET and Cre expression plasmid. These two are temp sensitive and will be lost at 37C. I tried to transform them into DH5-alpha, heat shock at 42C, then incubate at 30C for 1 hour, plate onto LB with Tc, and incubate at 30C for two nights. Cre plate did not grow. Red/ET plate had a few big colonies. I inoculated 3 colonies and miniprep DNA, but got very low concentration. Anyone has similar experience? Should I incubate them at 30C longer after heat shock? Why the conc. is so low?Anyone has the sequence and map information about these two plasmid? Thanks. pls reply here or e-mail to hzs108@gmail.com


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frasermoss
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Posted By frasermoss on 5/1/2009 18:04 PM
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The maps and sequences are freely available at the Gene Bridges web site

Here is the link to the manual for the Quick & Easy BAC Modification Kit

You should download the specific manual for the product that you have here www.genebridges.com/gb/products.php

 

 


Last edited May 01, 2009, 20:05 PM by frasermoss



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