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Latest revision as of 12:09, 25 February 2015
Cloning of 1002-pLysN
[edit]Purpose: Cloning of 1002-pLysN by PCR
Date: 2012-2-10--2012-2-17
Notebook:003737
Page:
Clone ID:1002
Experimenters: yingying
Results
[edit]cloning
[edit]Date:2012-2-10
1. PCR LacZProm-Sal1-F + LacZProm-R using pUC19 as template. 15 cycles
2. PCR T7LysN-F + T7LysN-BamH1-R, using pLysS as template, 15 cycles
pLysS plasmid can be extracted from BL21(DE3) pLysS
3. DO not run gel, Mix two PCR product, Purify PCR product using column, elute with 20 ul water.
4. PCR using LacZProm-Sal1-F + T7LysN-BamH1-R using step 3 eluate as template (all ~ 20 ul),25 cycles
| band 1 |
band 2 | |
| PCR product |
pACYC184 1003 | |
| digest enzymes |
Sal1 + Bamh1 | |
Colony Screening
[edit]Date: 2012-2-17
Make miniprep and digest with Sal1 + Bamh1
clone 1,2,3,5,6 may be right, need to sequence
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
<br>
Cloning of 1002-pLysN
[edit]Purpose: Cloning of 1002-pLysN by PCR
Date: 2012-2-22--2012-2-27
Notebook:003737
Page:
Clone ID:1002
Experimenters: yingying
Results
[edit]cloning
[edit]Date:2012-2-10
1. PCR LacZProm-Sal1-F + LacZProm-R using pUC19 as template. 15 cycles
2. PCR T7LysN-F + T7LysN-BamH1-R, using pLysS as template, 15 cycles
pLysS plasmid can be extracted from BL21(DE3) pLysS
3. DO not run gel, Mix two PCR product, Purify PCR product using column, elute with 20 ul water.
4. PCR using LacZProm-Sal1-F + T7LysN-BamH1-R using step 3 eluate as template (all ~ 20 ul),25 cycles
| band 1 |
band 2 | |
| PCR product |
pACYC184 1003 | |
| digest enzymes |
Sal1 + Bamh1 | |
Colony Screening
[edit]Date: 2012-2-24
Make miniprep and digest with Sal1 + Bamh1
clone 1,2,3,5,6 may be right, need to sequence
Result intepretation/Summary
[edit]- <br>