Yijia LI 7 23 2018
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Date:7/23/2018
[edit]Miniprep of 931_3206_129
Date:7/24/2018
[edit]931 subclone
Date:7/25/2918
[edit]3218 miniprep digestion
Date:7/26/2918
[edit]3206 PCR screening 1
PCR use 2x homemade Taq mix
3206 PCR screening 2
PCR use 2x homemade Taq mix
0.1Taq QC test
| #Lane | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
| Name | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 0.1Taq | 2X BestTaq Mastermix | Excellgen |
| Concentration | 880ng/ul | 220ng/ul | 110ng/ul | 55ng/ul | 27.5ng/ul | 13.75 | 6.875 | 3.43 | 1.71 | 0.85 | + control | +control |
miniprep digestion 3225_999_931
| # | Lane | Vector | Enyzmes | Size | note |
| 3225 | 1-4; 5 is + control | 1851 pT7A3-GST | Nhe1+Xho1 | 383bp | |
| 999 | 6-9;10 is +control | 1873 | Nde1+Xho1 | 1182bp | |
| 931 | 11-14;15 is + control | 931-1 | HindIII+Xho1 | failed |
Miniprep digestion of 941_3244_998
| # | Lane | Vector | Method | Enzymes | Size | Note |
| 941 | 1-4; 5 is + control | 947 | Subclone | Nde1+Xho1 | 456bp | |
| 3244 | 6-9 | 3242 | PCR cloning | Nde1+HindIII | 739bp | vector,wrong |
| 998 | 10-13 | 1708 | Gibson Assembly | HindIII+Xho1 |
Date:7/27/2018
[edit]2nd miniprep digestion_3225_999_998
| # | Lane | Enzymes | Vector | Size | Note |
| 3225 | 1-4 | Nco1+Xho1 | 1851 | three fragment.5288bp;738bp;361bp | #3,#4 |
| 999 | 5-8 | HindIII+Xho1 | 1873 | 1115bp | |
| 998 | 9-12 | HindIII+Kpn1 | 1708 | should have four fragments | Failed |
miniprep digestion_3249_3206_931-1
| # | Lane | Vector | Enyzmes | Size | note |
| 3249 | 1-4; 5 is + control;6 is vector - control | 1834 | Sph1+Xho1 | 1156bp | |
| 3206 | 7-10 | 3165 | BamH1+HindIII | Failed | |
| 931-1 | 11-14; 15 is +control, 16 is - control | 927 | Nde1+HindIII | ~989bp | lane 11,lane 12 |
