3249 pT7A3-GST-Avitag-BirA
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Cloning of 3249 pT7A3-GST-Avitag-BirA
[edit]Purpose: Cloning of 3249 pT7A3-GST-Avitag-BirA
Date: 07, 25, 2018
Notebook:
Page:
Clone ID: 3249
Experimenters: yijia
Results
[edit]Cloning
[edit]insert digestion
Vector digestion
Vector is 1834
1st digestion
2nd digestion
Colony screen
[edit]miniprep digestion_3249_3206_931-1
| # | Lane | Vector | Enyzmes | Size | note |
| 3249 | 1-4; 5 is + control;6 is vector - control | 1834 | Sph1+Xho1 | 1156bp | |
| 3206 | 7-10 | 3165 | BamH1+HindIII | Failed | |
| 931-1 | 11-14; 15 is +control, 16 is - control | 927 | Nde1+HindIII | ~989bp | lane 11,lane 12 |
10/27/2018
3274 is no good, multiple bands after NcoI-Nde1 digestion, or NdeI digestion,
so to check parent vector 3249, digest with NcoI and Nde1--> multiple bands
10/27/2018
retransform NEBTurbo, make miniprep, digestion
Looks like there are multiple Nde1 sites, need to sequence it.
sequencing:
3249 # 3 is good.
Nde1 cut--> multiple bands could be due to contamination of Nde1?
