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

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Autocreate 2013/11/13 Entry for User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli)
(Date)
Line 9: Line 9:
=Date=
=Date=
-
'''List title'''
+
'''Troubleshooting BD002 PCR Linearization for GG Assembly'''
-
# List items
+
 
 +
 
 +
 
 +
{| {{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'''
 +
|-
 +
| 11/9/2013||9 ng Plasmid: BD002||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||||
 +
|-
 +
| 11/9/2013||9 ng Plasmid: KAH187 (CMV)||[95C (30 sec.), 45C (45 Sec.), 72C (6 min.)]||||
 +
|-
 +
| 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
 +
|-
 +
| 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).
 +
|-
 +
| 11/11/2013||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 
 +
|-
 +
| 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
 +
|-
 +
| 11/11/2013||0.09ng Plasmid: BD002||[95C (30 sec.), 55C (45 Sec.), 72C (6 min.)]||||Increase the annealing temperature to 55C
 +
|-
 +
|}

Revision as of 14:36, 13 November 2013

PcTF Subcloning in E. coli Main project page
Previous entry      Next entry

Date

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
Personal tools