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

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| 11/9/2013||9 ng Plasmid: KAH187 (CMV)||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||[[Image:20131109 123553 (3).jpg |250px]]||
| 11/9/2013||9 ng Plasmid: KAH187 (CMV)||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||[[Image:20131109 123553 (3).jpg |250px]]||
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|-
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| 11/10/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||||Gel Extract the band close to 5K bps No result
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| 11/10/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||[[Image:20131110 122904 (2).jpg |250px]]||Gel Extract the band close to 5K bps No result
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| 11/10/2013||0.09ng 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).
| 11/10/2013||0.09ng 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).

Revision as of 15:26, 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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