1504 pT7A3-T4DNA-Pol-Exo-

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Cloning of 1504 pT7A3-T4DNA-Pol-Exo-

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Purpose: Cloning of1504 pT7A3-T4DNA-Pol-Exo-==

Date: 2014-9----

Notebook:0

Page:

Clone ID:1504

Experimenters:  shanyi

Results

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CLONING

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PCR using 1765 pET28b-T4DNAPol as template

Fragment 1
T4DNAPol-1-1 CGACTAGCTAGCAAAGAATTTTATATCTCTATTGAAACAGTCGG
T4-p43D219A-R catgatatacggaacgGcaaacccctcaatattc

Fragment 2
T4-p43D219A-F gaatattgaggggttt gCc gttccgtatatcatg
T4DNAPol-3-24 CGATCGGCGGCCGCTCAGCCAAACAGGAAGTCTAACGAAGCTTTTTCTTCATA


join two fragments by PCR


digest with NheI+NotI


Colony Screening

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Date: 2014_8

Make miniprep, use  HindIII to find positive clone




Result intepretation/Summary

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Conclusion

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