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Working backwards from a construct?


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#1 BiochemistryRA

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Posted 11 August 2014 - 05:55 AM

So I have a construct created with my gene insert and vector. I tried to express it in DE3 cells but it wouldn't work, I think the issue is that the insert is too big.

Just wondering if there is a way to go back from my construct, take out the gene, shorten it and start again? I have to work backwards from the construct.

Thank you in advance!



#2 bob1

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Posted 11 August 2014 - 01:25 PM

What do you mean too big an insert? Size of insert is unlikely to be the problem, it is far more likely that there is either a problem with the plasmid itself (insert not in frame, promoter missing), or that the product is toxic, or that the cells aren't inducing properly.






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