938 pET30b-PNGase F
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 |
subclone of 938
Size:1145bp
V:1987 pT7A3
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
938 miniprep test
1:938-1
2:938pJet1.2
Enzymes:EcoRV+XhoI
Size:552bp
3:938-1
4:938pJet1.2
Enzymes:NdeI+XhoI
Size:1071bp
Date:4/5/2018
[edit]938 subclone
Size:1145bp
Insert:938pJet1.2
Vector:1727 pT7A30
Enzymes:Nde1+Xho1
Miniprep digestion of 938_939_984
| # | Lane | Enzymes | Size | Note |
| 938 | 1-4;5 is + control(sequenced 938pjet1.2) | Nde1+Xho1 | 1145bp | good |
| 939 | 6-9 ; 10 is + control(sequenced 939pJet1.2) | Nde1+Xho1 | 780bp | good |
| 984 | 11-14; 15 is + control(sequenced 982) | Xba1+Nhe1 | 957bp | need to redo |
