3122 pET30b-CRM197
in pJet1.2
--Paul.xu (talk) 09:04, 21 December 2017 (EST)
12/27/2017
938 digestion with Nde1-Xho1, Fastdigest Buffer
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8,9 | 10,11 | 13 |
| 938 PNGase_F | 939 Tte-ExoIII |
942 VVN |
3122 CRM197 |
931, 1, PAK16D ligase |
931, 2, PAK16D ligase |
1855 yPolyA-Pol |
1540 | 2046 | 2046 |
| Nde1-Xho1 | Nde1-Xho1 |
Nco1-Xho1 |
Nde1-Xho1 |
Nde1-HindIII |
HindIII-Xho1 | Nde1-Xho1 | Nde1-Xho1 | Nde1-Xho1 | Nco1-Xho1 |
| pJet1.2 | pJet1.2 |
pJet1.2 |
pJet1.2 |
pJet1.2 |
pJet1.2 |
pEC2- |
pET30b | pET28b | pET28b |
| 1145 |
780 |
716 |
1798 |
1174 |
986 |
1713 | there is no Xho1 site | 1377 | ~1377 |
Date:3/30/2018
[edit]Miniprep digestion of 3122_938_939
3122:lane 1-4 lane 5 :+ size:1743 bp
938: lane 6-9 lane 10:+ size:1145 bp
939: lane 11-14 lane 15:+ size:780 bp
Positive control are insert-pjet1.2
The small band size of vector 1727:193 bp
redo Miniprep digestion of 3122_938_939
1:3122 size:1743bp
2:3122pjet1.2 sequenced +
3:938 size:1145 bp
4:938pjet1.2 sequenced +
5,6,7:939 size:780 bp
8:939pjet1.2 sequenced +
Enzymes:NdeI+XhoI
Date:4/5/2018
[edit]824subclone
Insert size:1743bp
Vector:1987pT7A3
Date:5/29/2018
[edit]Subclone and Vector digestion
3122: what is the vector name? clone #500 , insert size should be 2055 (Nde1 + Xho1)
Miniprep digestion of 2503_3122_3197_3191
| # | lane | Enzymes | Size | Note |
| 2503 | 1-4 | Nde1+Xho1 | 951bp | #1,2 seems good |
| 3122 | 5-8 | Nde1+Xho1 | 1743bp | #5,6,8 seems good |
| 3197 | 9-12 | BamH1+Nco1 | 1 band | may not good |
| 3191 | 13-16 | BamH1+Nco1 | 722bp | #14,15,16 seems good |
3122 could be parental vector
Miniprep of 3122_3210
Miniprep of 3122
