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

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Date

Troubleshooting BD002 PCR Linearization for GG Assembly


Date Plasmid Conc. Therm Cycling Conditions Gel Picture Comment
11/9/2013 9 ng Plasmid: BD002 [95C (30 sec.), 45C (45 Sec.), 72C (6 min.)] 20131109 123553 (2).jpg
11/9/2013 9 ng Plasmid: KAH187 (CMV) [95C (30 sec.), 45C (45 Sec.), 72C (6 min.)] 20131109 123553 (3).jpg
11/10/2013 0.09ng Plasmid: BD002 [95C (30 sec.), 45C (45 Sec.), 72C (6 min.)] 20131110 122904 (2).jpg Gel Extract the band close to 5K bps No result
11/10/2013 0.09ng Plasmid: BD002 [95C (30 sec.), 45C (45 Sec.), 72C (6 min.)] 20131110 122904 (3).jpg Amplified with V0200 Sequencing Primers to make sure Kozak-HSV-CFP-NLS-Stop-PolyA (2K bps are there).
11/11/2013 Plasmid: BD002 [95C (30 sec.), 49C (45 Sec.), 72C (6 min.)] 20131110 213024 (2).jpg 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 0.09ng Plasmid: BD002 [95C (30 sec.), 49C (45 Sec.), 75C (6 min.)] 20131110 213024 (3).jpg Increase the extention temperature to 75C and annealing temperature to 49C
11/11/2013 0.09ng Plasmid: BD002 [95C (30 sec.), 55C (45 Sec.), 72C (6 min.)] 20131111 095957 (2).jpg Increase the annealing temperature to 55C