User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/11/13

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| 11/11/2013||Plasmid: BD002||[95C (30 sec.), 49C (45 Sec.), 72C (6 min.)]||[[Image:20131110 213024 (2).jpg |250px]]||Column 1,2: 9 ng Plasmid, Column 3,4: 0.09 ng Plasmid, Column 5,6: 0.5 ml primer from 10mM solution   
| 11/11/2013||Plasmid: BD002||[95C (30 sec.), 49C (45 Sec.), 72C (6 min.)]||[[Image:20131110 213024 (2).jpg |250px]]||Column 1,2: 9 ng Plasmid, Column 3,4: 0.09 ng Plasmid, Column 5,6: 0.5 ml primer from 10mM solution   
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| 11/11/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 49C (45 Sec.), 75C (6 min.)]||||Increase the extention temperature to 75C and annealing temperature to 49C
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| 11/11/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 49C (45 Sec.), 75C (6 min.)]||[[Image:20131110 213024 (3).jpg |250px]]||Increase the extention temperature to 75C and annealing temperature to 49C
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| 11/11/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 55C (45 Sec.), 72C (6 min.)]||||Increase the annealing temperature to 55C
| 11/11/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 55C (45 Sec.), 72C (6 min.)]||||Increase the annealing temperature to 55C
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Revision as of 15:38, 13 November 2013

PcTF Subcloning in E. coli Main project page
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Troubleshooting BD002 PCR Linearization for GG Assembly


Date Plasmid Conc. Therm Cycling Conditions Gel Picture Comment
11/9/20139 ng Plasmid: BD002[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]
11/9/20139 ng Plasmid: KAH187 (CMV)[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]
11/10/20130.09ng Plasmid: BD002[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]Gel Extract the band close to 5K bps No result
11/10/20130.09ng Plasmid: BD002[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]Amplified with V0200 Sequencing Primers to make sure Kozak-HSV-CFP-NLS-Stop-PolyA (2K bps are there).
11/11/2013Plasmid: BD002[95C (30 sec.), 49C (45 Sec.), 72C (6 min.)]Column 1,2: 9 ng Plasmid, Column 3,4: 0.09 ng Plasmid, Column 5,6: 0.5 ml primer from 10mM solution
11/11/20130.09ng Plasmid: BD002[95C (30 sec.), 49C (45 Sec.), 75C (6 min.)]Increase the extention temperature to 75C and annealing temperature to 49C
11/11/20130.09ng Plasmid: BD002[95C (30 sec.), 55C (45 Sec.), 72C (6 min.)]Increase the annealing temperature to 55C
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