2213 pET30b-Dest-C-Avi-His

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Cloning of 2213 pET30b-Dest-C-Avi-His

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Purpose: Cloning of2213 pET30b-Dest-C-Avi-His==

Date: 2014-8----

Notebook:0

Page:

Clone ID:2213

Experimenters:  shanyi

Results

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CLONING

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PCR gateway cassette from 1336

1 ul of 10 uM attR1-Fix-Nde1-F tagcac cat atg gaa TCAACAAGTTTGTACAAAAAAG
1 ul of 1 uM attR2-AviTag-R2 ( ctttcttgtacaaagtggt GGCGGCCTGAACGACATCTTCGAGGCCCAGAAAATCGAGTGGCAC) GTGCCACTCGATTTTCTGGGCCTCGAAGATGTCGTTCAGGCCGCCaccactttgtacaagaaag

1 ul of 10 uM attR2-AviTag8H-Kpn1-EcoR1-R1 (CAGAAAATCGAGTGGCACGAG CAC CAC CAT CAT CAC CAT CAC CAC TGA GGTACC gaattc tacga) tcgtagaattcGGTACCTCAGTGGTGATGGTGATGATGGTGGTGCTCGTGCCACTCGATTTTCTG


digest with Nde1 + EcoR1, clone into Nde1 + EcoR1 digested vector 16 pET30b.Amp,




Colony Screening

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Date: 2014_7

Make miniprep, use Nde1 + EcoR1to find positive clone


Result intepretation/Summary

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Conclusion

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