Yijia LI 4 9 2018
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Date:4/9/2018
[edit]1.Pick up clonies
2.collect cells
3.redo subclone
Date:4/10/2018
[edit]Miniprep digestion of 3166pMiniT2_944pMiniT2_3127pMiniT2
Insert size:
3166pMiniT2:1234bp SalI+BsrGI
944pMiniT2 :2334bp NdeI+XhoI
3127pMiniT2:402bp Nde1-HindIII
944 subclone
3166 subclone
Miniprep digestion of 945_986_944_3127
| # | Lane | Size | Enzymes | Note |
| 945 | 1-4 | 1326bp | Xba1-Nco1 |
subclone |
| 986 | 5-8 | 774bp | HindIII+XhoI | ~4kbp |
| 944 | 9-12 | 2334bp | NdeI+XhoI | PCR |
| 3127 | 13-16 | 402bp | Nde1-HindIII |
Gibson Mix |
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 |
Date:4/11/2018
[edit]Miniprep digestion of 3152_985
| # | Lane | Size | Enzymes | Note |
| 3152 | 1-4 | 371bp | BsrG1-Nhe1 |
size is the same as digested vector,need to do sequencing |
| 985 | 5-8, Lane 9 is +control | 402bp | Nde1+Kpn1 | good |
Date:4/12/12018
[edit]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 |
Date:4/13/2018
[edit]Miniprep of 944_3166_3127
| # | Lane | Size | Enzymes | Result | Note |
| 944 | 1-4; 5 is 944pminiT2 | 2334bp | Nde1+Xho1 | Size is a little bigger than 2334 bp, need to do sequencing | 944pMiniT2 is from PCR |
| 3166 | 5-9; 10 is 3166pminiT2 | 1234bp | BsrG1+Sal1 | to do sequence | 3166pMiniT2 is from PCR |
| 3127 | 11-14; 14 is 3127pMiniT2 | 402bp | Nde1+HindIII | to do sequence | 3127pMiniT2 is from PCR |
Date:4/14/2018
[edit]Miniprep of 989_990_991_992
Miniprep of 993_995
Miniprep of 986
#Ends Coordinates Length (bp)
1 HindIII-HindIII 1124-3468 2345
2 HindIII-HindIII 93-1123 1031
3 HindIII-HindIII 3469-92 893
