942 pET28b-pMBP-VVN
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:4/5/2018
[edit]subclone of 942
Size:716bp
Vector:947 pT7A3
Insert:942
Miniprep digestion of 984_942_824_3166
| # | Lane | Size | Enzymes | Note |
| 984 | 1-4 | Xba1+NdeI | Subclone | |
| 942 | 5-8 | 716bp | Nco1-Xho1 |
|
| 824 | 9-12 | 969 |
Nde1 + Xho1 |
|
| 3166 | 13-16 | 1234bp | Sal1+BsrG1 |
PCR |
test for selecting correct subclone of 986_944_945_942_824
| # | Lane | control | Size | Enzymes | Note |
| 986 | 1-2 | Lane 3 1708 vector | HindIII+XhoI |
undigested vector | |
| 944 | 4-6 | Lane 7 944pMIniT2 | 2334bp | NdeI+XhoI |
from PCR, No.4 might be right one |
| 945 | 8 | Lane 9 945pMiniT2 | 1326bp | Xba1-Nco1 |
good |
| 942 | 10-12 | Lane 13 942pJet1.2 | 716bp | Nco1-Xho1 |
good |
| 824 | 14-15 | Lane 16 832pMiniT2 | 969bp | Nde1 + Xho1 |
