129 pL42-TthMuts
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Cloning of 129 pT7A30-TthMuts
[edit]Purpose: Cloning of 129 pL42-TthMuts
Date: 06, 27, 2018
Notebook:
Page:
Clone ID: 129
Experimenters: yijia
Results
[edit]Cloning
[edit]PCR from 212 pL6H-TthMuts
PCR of 129
redo PCR
Template:212
commerical 2xPCR mix,5x enhancer
Date:8/4/2018
[edit]redo PCR
Vector digestion
Vector is 2289
Colony screen
[edit]Miniprep of 129_994_1000_3206
| # | Vector | Lane | Enzymes | Size | Clone# | Note |
| 129 | 1801 | 1-4 | Nde1+HindIII | 2433 | 2,4 look good | 129-2;129-4 |
| 994 | 3225 | 5-8 | BamH1+Xho1 | 609 | 5,6,look good | Vecotor doesn't have BamH1 Site,994-1;994-2 |
| 1000 | 1708 | 9-12 | HIndIII+Xho1 | 118 | 11-13 good | |
| 3206 | 3165 | 13-16 | HIndIII+BamH1 | 1460 | need to do PCR screening |
wrong vector
Date:7/26/2018
129 PCR screening
Date:8/7/2018
[edit]129 PCR Screening
Primers:T7-R+212-F
Result:#1-7 are good, grow up in kana
Miniprep digestion of 129