998 pT7A3-6His-NLS-dCas9-NLS-6xHis
Vector digestion
998 Vector was not digested completely
PCR of 998_1000
Miniprep digestion of 998_1000_994_995
| # | Lane | Enzymes | Size | Note |
| 998 | 1-4 | HindIII+Xho1 | ||
| 1000 | 5-8 | HIndIII+Xho1 | ||
| 994 | 9-12 | BamH1+Xho1 | ||
| 995 | 13-16 | BamH1+Xho1 |
Cloning of 998 pT7A3-6His-NLS-dCas9-NLS-6xHis
[edit]Purpose: Cloning of 998 pT7A3-6His-NLS-dCas9-NLS-6xHis
Date: 06, 27, 2018
Notebook: 008840
Page:008958
Clone ID: 998
Experimenters: yijia
Cloning
[edit]998 PCR
Miniprep of 3206_995_998
Miniprep digestion of 3218_998
| # | lane | Enzymes | Size | Result |
| 998 | 9-12 | HindIII | ||
| 13-16 | HindIII+Xho1 |
redo
[edit]998 PCR
| # | Lane | Enzymes | Buffer |
| Kappa | 1,2 |
Kappa HiFi |
5xKappa buffer |
| Takara | 3,4 | Takara DNA polymerase | 5xTakara buffer |
| Q5 | 5,6 | Q5 HiFi | 5xphusiob HF hot start buffer |
redo PCR
Date:8/2/2018
[edit]vector digestion of 3206_998
Results
[edit]Colony screen
[edit]Date:8/7/2018
[edit]Miniprep digestion of 998_931-1_931-1pjet1.2_129
Miniprep digestion 998
1-3:HindIII+Xho1
# Ends Length (bp)
1 HindIII-HindIII 2345
2 HindIII-HindIII 1034
3 HindIII-XhoI 774
4 XhoI-HindIII 118
4-6:Nhe1+Kpn1 679bp
Date:8/8/2018
[edit]miniprep digstion 998
choose #1,#3 to do another digestion
Miniprep digestion of #1,#3
Date:8/13/2018
[edit]redo vector digstion
998 PCR Screening
998 miniprep 1
998 Miniprep 2
