Yijia LI 4 2 2018
Date:4/2/2018
[edit]subclone of 984
Vector:1987
Insert:982
Size:957 bp
Enzymes:Xba1-Nhe1
redo subclone of 3152
Vector:3163
Insert: 3009pMiniT
enzymes:BsrGI+NheI
redo Subclone of 947
Vector:1727
Insert:947pMiniT
Size:1344 bp
Date:4/3/2018
[edit]redo PCR of 3127
new primer
3127-HindIII-R
3127-NheI-F2
size:403bp
extension time:30s
PCR of 986
F1:986-F1+986-R1
F2:986-F2+986-R2
Size:4kbp
Extension time:4min
Date:4/4/2018
[edit]miniprep digestion of 984_947_3152
- Name Enzymes Size:
1-4:984 XbaI+NheI 957bp
5 :982(973-1pMiniT) XbaI+NheI
6-9:947 XbaI+NcoI-HF 1344bp
10:947pMiniT XbaI+NcoI-HF
11:947(from PCR cloning) XbaI+NcoI-HF
12-15:3152 BsrGI+NheI 371bp
16: 3162(Vector) BsrGI+NheI
3122 miniprep test digestion
1:3122-2
2:3122pJet1.2
enzymes:NdeI+SalI 265bp
3:3122-2
4:3122pJet1.2
Enzymes:EcoRV+SalI
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
939 miniprep test digestion
1:939-1 1-4 enzymes:NdeI+AgeI
2:939-2 1-4 Size: 421bp
3:939-4
4:939pJet1.2
5:939-1 5-8 enzymes:XhoI+AgeI
6:939-2 5-8 Size:355bp
7:939-4
8:939pJet1.2 Results:all of them have bands
Date:4/5/2018
[edit]subclone of 942
Size:716bp
Vector:947 pT7A3
Insert:942
subclone of 986
Vector:1708
For 986 cloning
Subclone of 931
Vector:1987pT7A3
Insert:
F1:1174 bp
F2:986 bp
824subclone
Size:~969bp
vector:1987
Insert:832pMiniT
824subclone
Insert size:1743bp
Vector:1987pT7A3
Subclone 984
Vector:1987
Insert size:957 bp
Vector digestion for 3127
PCR of 944 F1+F2
Template:367, cut out band then combine
new way of 986 PCR
redo subclone of 3152
Date:4/6/2018
[edit]939 suclone
Size:780bp
Insert:939pJet1.2
Vector:1727 pT7A30
Enzymes:Nde1+Xho1
938 subclone
Size:1145bp
Insert:938pJet1.2
Vector:1727 pT7A30
Enzymes:Nde1+Xho1
3122 subclone
Size:1743bp
Insert:3122pJet1.2
Vector:1727 pT7A30
Enzymes:Nde1+Xho1
3127 subclone
Size:408 bp
Insert:939pJet1.2
Vector:1727 pT7A30
Enzymes:Nde1+HindIII
986 PCR
986 digestion as vector
Enzymes:HindIII+Xho1
945 vector diegstion
Enzymes:Xba1+Nco1
3166 PCR
Primer:3166-F+3166-R
Template:3017 pCEH0
Extension time:1min 30s
Size:1234bp
3152 subclone
| Lane# | Name | Note |
| 1 | Undigested vector | - control |
| 2 | digested vector | - control |
| 3 | digested vector---3016 pCEH0 | for subclone |
| 4 | digested insert---3009pMiniT2 | for subclone |
3129 digestion as vector
Enzymes:Sal1+BsrG1
945 subclone
Size:1326 bp
Enzymes:Xba1+Nco1
3165 PCR
944 PCR
Size:2334bp. might be wrong, because two bands in the gel.
931 subclone
Vector:1518 pET28b digested with Nde1+Xho1
F1:1174 bp, digested with Nde1+HindIII
F2:986 bp, digested with HindIII+ Xho1
985 subclone
Vector:1987 pT7A3 digested with' Nde1-Xho1 Insert:983(973-2 pMiniT2) digested with Nde1+Sal1
Size:500 bp
