Editing 1106 pFastBacDual2-TagGFP

Cloning of pFastBacDual2-TagGFP(1106)<br>Purpose: to clone pFastBacDual2-TagGFP<br>Date: 2012-7-18 to 2012-7-<br>Notebook: Xiaojing Wang<br>Page: 005300 

Clone ID: 1106<br>Experimenters: Xiaojing Wang 

Results<br> 

digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed<br> 

FastBac3-T + FastBac3-B <br> 

[[Image:1106 v digest1 7.18.2012.JPG]][[Image:1106 v digest2 7.19.2012.JPG]][[Image:1106 v digest3 7.19.2012.JPG]] 

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Transform DH5alpha 

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== '''Cloning of 1106 pFastBacDual2-TagGFP'''  ==

'''Purpose:''' Cloning of 1106 pFastBacDual2-TagGFP 

'''Date: '''July 26, 2012<br> 

'''Notebook:&nbsp;''' 

'''Page:''' 

'''Clone ID: '''1106 

'''Experimenters: '''Jennifer Truong 

== '''Results'''  ==

=== Cloning  ===

Date:&nbsp;July 26, 2012<br> 

Digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed <br> 

FastBac3-T + FastBac3-B<br>(BamH1) Nde1 T7 promoter + RBS BamH1 Nco1 EcoR1 <br>GATCATATGTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCGAAGGAGATATAGGatccatgg<br> TATACATTATGCTGAGTGATATCCCCTTAACACTCGCCTATTGTTAAGCTTCCTCTATATcctaggtaccTTAA 

FastBac3-T<br>GATCATATGTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCGAAGGAGATATAGGatccatgg<br> FastBac3-B<br>AATTccatggatccTATATCTCCTTCGAATTGTTATCCGCTCACAATTCCCCTATAGTGAGTCGTATTACATAT<br> 

[[Image:1106 Vector.JPG]]<br> 

Digest 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, see lane one, cut out circled band, purified and ligated (20 uL&nbsp;vector, 10 uL&nbsp;annealed oligo, 3 uL&nbsp;ligation buffer, 1 uL&nbsp;T4 Ligase) overnight, transform tomorrow<br> 

=== <u></u>Colony Screening<br>  ===

Date:&nbsp;July 30, 2012<br> 

[[Image:1106 Screening.JPG]]<br> 

Digested MiniPrep with RE BamH1 + EcoR1, expected size ~6000 bp<br> 

Samples 1, 3, 4, 6, 7 may be correct 

[[Image:1106 Digestion.JPG]] 

Digested MiniPrep with RE Nde1 and Kpn1, expected size 1053 bp 

All negative? 

== '''Result intepretation/Summary'''  ==

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== '''Conclusion'''  ==

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== '''Cloning of 1106-pFastBacDual2-TagGFP'''  ==

'''Purpose:''' Cloning of 1106-pFastBacDual2-TagGFP 

'''Date: '''2012-9-25---2012-9-27

'''Notebook:0''' 

'''Page:''' 

'''Clone ID:1106''' 

'''Experimenters:''' yingying 

== '''Results'''  ==

=== cloning  ===

[[Image:559+567+1352+1248+1106 clone 9-25-2012.JPG]]<br> 

digest (lane7) 1105 pFastBacDual-TagGFP with BamH1 + EcoR1, ligate to annealed FastBac3-T + FastBac3-B<br> 

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=== Colony Screening  ===

Date: 2012-9-27

mini prep and digest with Nde1 + Kpn1<br>

[[Image:1106+1402_digest_detect_9-27-2012.JPG]]

clone 1,2,3,4,5,6 may be right

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== '''Result intepretation/Summary'''  ==

*&amp;amp;lt;br&amp;amp;gt;

== '''Conclusion'''  ==

&amp;amp;lt;br&amp;amp;gt; 

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&amp; 

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[[Category:Cloning]]