2436-pET30-30GFP: Difference between revisions

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Latest revision as of 16:23, 4 May 2015

Cloning of 2436-pET30-30GFP

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Date:  2015-April

Clone ID: 2436

Experiment:  Xin

Results:

1. Gene Synthesis by two sections:

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Section I: 1-1 __ 1-10; Section II: 1-11 __1-20

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combine two sections together, PCR again.

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  2. Clone:

Insert the PCR product into vector 440.

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Ligation and transformation. 

Screening:

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 RTENOTITLE  The insectionthe size is about 1200bp, but should be about 850 bp.

Redigested again using different enzyme: XhoI/ NdeI
RTENOTITLE  The size is still not right.

Selected one clony to sequence, the clone looks all right. The marker on the argrose gel isn't very accurcy.

Fix on April, 28, 2015

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 First PCR fragment: use the oligos 2436-30-GFP-1;2436-30-GFP-12 as primers

 Second PCR fragment: use the oligos 2436-30-GFP-13;2436-30-GFP-20 as primers

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Join two PCR together:
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Screening:
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All 8 colonies have right insert.

Result intepretation/Summary

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Conclusion

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Result intepretation/Summary

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Conclusion

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