User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/11/13: Difference between revisions
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''' | '''Troubleshooting BD002 PCR Linearization for GG Assembly''' | ||
# | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''Date''' | |||
| align="center" style="background:#f0f0f0;"|'''Plasmid Conc.''' | |||
| align="center" style="background:#f0f0f0;"|'''Therm Cycling Conditions''' | |||
| align="center" style="background:#f0f0f0;"|'''Gel Picture''' | |||
| align="center" style="background:#f0f0f0;"|'''Comment''' | |||
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| 11/9/2013||9 ng Plasmid: BD002||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]|||| | |||
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| 11/9/2013||9 ng Plasmid: KAH187 (CMV)||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]|||| | |||
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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.)]||||Amplified with V0200 Sequencing Primers to make sure Kozak-HSV-CFP-NLS-Stop-PolyA (2K bps are there). | |||
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| 11/11/2013||Plasmid: 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 | |||
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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.), 55C (45 Sec.), 72C (6 min.)]||||Increase the annealing temperature to 55C | |||
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Revision as of 12:36, 13 November 2013
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DateTroubleshooting BD002 PCR Linearization for GG Assembly
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