1780-pT7A3-T7-RNAPol-Y639F

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Cloning of 1780-pT7A3-T7-RNAPol-Y639F

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Purpose: Cloning of 1780-pT7A3-T7-RNAPol-Y639F

Date: 2013-7-16---2013-7-23

Notebook:0

Page:

Clone ID:1780

Experimenters: yingying

Results

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cloning

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1. PCR using BL21(DE3) genomic DNA as template

T7RNAPol-Nco1-F2
T7RNAPol-Y639F-R


2. PCR using 1080 pBad24-RNAPol
T7RNAPol-Y639F-F
T7RNAPol-Xho1-R2


3. Join two fragments using
T7RNAPol-Nco1-F2
T7RNAPol-Xho1-R2


lane
 1
 2

 PCR product
 1279 pT7A3
enzymes
    Nco1 + Xho1


Colony Screening

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Date: 2013-7-22

Make miniprep and digest with Nco1 + Xho1


2013-7-23

digest with Nco1+Kpn1


Result intepretation/Summary

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Conclusion

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