1048-pET30b-RNaseI

Cloning of 1048-pET30b-RNaseI

Purpose: Cloning of1048-pET30b-RNaseI

Date: 2012-3-27--2012-3-30

Notebook:

Page:003783

Clone ID:1048

Experimenters: yingying

Results

cloning

Date:2012-3-27

Fragment1: RnaseI-1 ~RnaseI-20, ~650 bp,gene synthesis
Fragment 2: RnaseI-21~40, ~740 bp,gene synthesis
PCR 2 fragments and then using Gibson mix to join them

digest :

	band 3	band 4
	Gibson product	pET30b
enzymes	Nde1+ Xho1

Colony Screening

Date: 2012-3-26

PCR screen  with T7 F+RnaseI-40

no product

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Conclusion

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Cloning of 1048-pET30b-RNaseI

Purpose: Cloning of1048-pET30b-RNaseI

Date: 2012-3-30--2012-4-2

Notebook:

Page:003789

Clone ID:1048

Experimenters: yingying

Results

cloning

Date:2012-3-30

Fragment1: RnaseI-1 ~RnaseI-20, ~650 bp,gene synthesis
Fragment 2: RnaseI-21~40, ~740 bp,gene synthesis
PCR 2 fragments then 

Fragment1+Fragment 2 as template

RnaseI-1+RnaseI-40 as primers

	band 3	band 4
enzymes	Nde1+ Xho1

Colony Screening

Date: 2012-4-3

miniprep and digest with Nde1+ Xho1

no band

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Conclusion

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Cloning of 1048-pET30b-RNaseI

Purpose: Cloning of1048-pET30b-RNaseI

Date: 2012-4-4--2012-4-10

Notebook:

Page:

Clone ID:1048

Experimenters: yingying

Results

cloning

Date:2012-3-27

Fragment1: RnaseI-1 ~RnaseI-20, ~650 bp,gene synthesis
Fragment 2: RnaseI-21~40, ~740 bp,gene synthesis
PCR 2 fragments then

Fragment1+Fragment 2 as template

RnaseI-1+RnaseI-40 as primers

digest :

	band 5	band 6
	PCR product	pET30b
enzymes	Nco1+ Xho1

Colony Screening

Date: 2012-4-10

clone1,2,3,5,6,8 may be right, need to sequence

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Conclusion

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