1106 pFastBacDual2-TagGFP
Cloning of pFastBacDual2-TagGFP(1106)
Purpose: to clone pFastBacDual2-TagGFP
Date: 2012-7-18 to 2012-7-
Notebook: Xiaojing Wang
Page: 005300
Clone ID: 1106
Experimenters: Xiaojing Wang
Results
digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed
FastBac3-T + FastBac3-B
Transform DH5alpha
Cloning of 1106 pFastBacDual2-TagGFP
[edit]Purpose: Cloning of 1106 pFastBacDual2-TagGFP
Date: July 26, 2012
Notebook:
Page:
Clone ID: 1106
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 26, 2012
Digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed
FastBac3-T + FastBac3-B
(BamH1) Nde1 T7 promoter + RBS BamH1 Nco1 EcoR1
GATCATATGTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCGAAGGAGATATAGGatccatgg
TATACATTATGCTGAGTGATATCCCCTTAACACTCGCCTATTGTTAAGCTTCCTCTATATcctaggtaccTTAA
FastBac3-T
GATCATATGTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCGAAGGAGATATAGGatccatgg
FastBac3-B
AATTccatggatccTATATCTCCTTCGAATTGTTATCCGCTCACAATTCCCCTATAGTGAGTCGTATTACATAT
Digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, see lane one, cut out circled band, purified and ligated (20 uL vector, 10 uL annealed oligo, 3 uL ligation buffer, 1 uL T4 Ligase) overnight, transform tomorrow
Colony Screening
[edit]Date: July 30, 2012
Digested MiniPrep with RE BamH1 + EcoR1, expected size ~6000 bp
Samples 1, 3, 4, 6, 7 may be correct
Digested MiniPrep with RE Nde1 and Kpn1, expected size 1053 bp
All negative?
Result intepretation/Summary
[edit]
Conclusion
[edit]
Cloning of 1106-pFastBacDual2-TagGFP
[edit]Purpose: Cloning of 1106-pFastBacDual2-TagGFP
Date: 2012-9-25---2012-9-27
Notebook:0
Page:
Clone ID:1106
Experimenters: yingying
Results
[edit]cloning
[edit]
digest (lane7) 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed FastBac3-T + FastBac3-B
Colony Screening
[edit]Date: 2012-9-27
mini prep and digest with Nde1 + Kpn1
clone 1,2,3,4,5,6 may be right
Result intepretation/Summary
[edit]- <br>
Conclusion
[edit]<br>
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