1002-pLysN

Cloning of 1002-pLysN

Purpose: Cloning of 1002-pLysN by PCR

Date: 2012-2-10--2012-2-17

Notebook:003737

Page:

Clone ID:1002

Experimenters: yingying

Results

cloning

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

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

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Conclusion

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Cloning of 1002-pLysN

Purpose: Cloning of 1002-pLysN by PCR

Date: 2012-2-22--2012-2-27

Notebook:003737

Page:

Clone ID:1002

Experimenters: yingying

Results

cloning

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

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

• <br>