2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
[edit]Purpose: Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Date: 2014-2-3---2014-2-6
Notebook:0
Page:
Clone ID:2014
Experimenters: yingying
Results
[edit]cloning
[edit]
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| 5 |
6 | |
| 1955 |
2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGFP | |
| BamH1 + Xho1 | ||
| 1 |
2 | |
| pcr |
2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGF | |
| BamH1 + Xho1 | ||
Colony Screening
[edit]Date: 2014-2-6
Make miniprep and digest BamH1+xba1
c2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGF
Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
[edit]Purpose: Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Date: 2014-2-
Notebook:0
Page:
Clone ID:2014
Experimenters: shanyi
Results
[edit]cloning
[edit]
| 1955 |
2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGFP | |
| BamH1 + Xho1 | ||
Colony Screening
[edit]Date: 2014-2-6
Make miniprep
digest BamH1+Xho1
digest with XbaI
for 2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGF there are three xbaI at 630bp, 1985bp(cleavage affected by meth), 2518bp, so digesting with XbaI can get two bands
for 2014 there is only one XhaI at 1988bp(cleavage affected by meth)
SENT 2014-1 TO SEQUENCE WITH CMV-SEQ-F
Result intepretation/Summary
[edit]Conclusion
[edit]
Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
[edit]Purpose: Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Date: 2014-2-24
Notebook:0
Page:
Clone ID:2014
Experimenters: yingying
Results
[edit]cloning
[edit]
PCR using clone1955 as template
LoxP-TurboRFP-BamH1-F3
TurboGFP-HindIII-Sal1-R
| lane |
1 |
2 |
| pcr |
2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGFP | |
| BamH1 + HindIII | ||
Colony Screening
[edit]Date: 201
Make miniprep and digest with
Result intepretation/Summary
[edit]Conclusion
[edit]Cloning of 2029 pEC/PEC2-LoxP-TurboRFP-ODC-2Stops-LoxP
[edit]Purpose: Cloning of 2029pEC/PEC2-LoxP-TurboRFP-ODC-2Stops-LoxP
Date: 2014-3-7--2014-3-10
Notebook:0
Page:
Clone ID:2029
Experimenters: yingying
Results
[edit]cloning
[edit]
| lane |
1 |
2 |
3 |
4 |
| PCR |
1454pEC |
PCR |
2015pEC2 | |
| BamH1 + Xho1 | ||||
Colony Screening
[edit]Date: 2014-3-10
Make miniprep and digest with BamH1 + Xho1
1-4:pEC vector
5-8:pEC2 vector
Result intepretation/Summary
[edit]Conclusion
[edit]sub Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP
[edit]Purpose: Cloning of 2014 pCMV-LoxP-TurboRFP-ODC-2Stops-LoxP-TurboGFP
Date: 2014-3-11-2014-3-12
Notebook:0
Page:
Clone ID:2014
Experimenters: yingying
Results
[edit]cloning
[edit]
| lane |
1 |
2 |
| 2029pEC-LoxP-TurboRFP-ODC-2Stops-LoxP |
2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGF | |
| BamH1 + Xho1 | ||
Colony Screening
[edit]Date: 2014-3-12
Make miniprep and digest with
| lane |
1 |
2 |
3 |
| 2014-7 |
2014-7 |
2013,2013 pCMV-FRT-TurboRFP-ODC-2Stop-FRT-TurboGF | |
| BamH1+Xho1 |
BamH1+Xba1 |
BamH1+Xba1 |
2014-3-24
BamH1+Xho1
There is no control for this digestion, you should digest with BamH1 alone, Xho1 alone and no digestion for at least one clone. Clone 1, and 2, 4 are not right. Clone #3 also looks weird. You MUST run a few controls if an experiment does not look right in the future.
2014-4-2
2014 transform DH5a and NEBturbo ,miniprep and digest with BamH1+Xho1/BamH1/Xho1
| lane |
1 |
2 |
3 |
4 |
5 |
6 |
7clone1 |
8clone1 |
| DH5a |
NEBturbo |
clone1 | ||||||
| BamH1+Xho1 |
BamH1 |
Xho1 | ||||||
2014-clone 1 digest with BamH1+EcoR1/EcoR1+Xho1
| lane |
1 |
2 |
| 2014-clone 1 | ||
| BamH1+EcoR1 |
EcoR1+Xho1 | |