1144 pT7A3-MBP-(3C)-Bst-Exo-
Cloning of 1144 pT7A3-MBP-(3C)-Bst-Exo-
[edit]Purpose: Cloning of1144 pT7A3-MBP-(3C)-Bst-Exo-
Date: 2013-3-
Notebook:
Page:
Clone ID:1144
Experimenters: Shanyi
Results
[edit]cloning
[edit]Date:2013-3-
Gene synthesis 3 PCR fragments
then use Fragment1,2,3 as template
Bst-Exo-1-1+Bst-Exo-3-18 as primers
digest :
| band 2,3 |
band 4 | |
| PCR product |
1514 | |
| enzymes |
Nde1+ Xho1 | |
Colony Screening
[edit]Date: 2013
SEND 1144-3 TO SEQUENCE, MANY GAPS
Cloning of 1144 pEC-Bst-Exo-
[edit]Purpose: Cloning of1144 pEC-Bst-Exo-
Date: 2013-3-
Notebook:
Page:
Clone ID:1144
Experimenters: Shanyi
Results
[edit]cloning
[edit]Date:2013-3-
Gene synthesis 3 PCR fragments
then use Fragment1,2,3 as template
Bst-Exo-1-1+Bst-Exo-3-18 as primers
digest :
| band 1 |
band 3 | |
| PCR product |
1454 | |
| enzymes |
Nde1+ Xho1 | |
Colony Screening
[edit]Date: 2013
DIGEST WITH NdeI , XhoI and BamHI (should get there bands=>350bp, 1500bp,2500bp)
SEND 1144-1 TO SEQUENCE, missing 3nt
send 1144-2 to sequence, missing 1G(SEQUENCE FROM T7 TERM AND M13R)
cloning
[edit]Date:2013-3-
Fragment 1, use Bst-Exo-1-1 and Bst-Exo-3-12
Fragment 2,use Bst-Exo-3-13 and Bst-Exo-3-18
then use Fragment 1and2 as template
Bst-Exo-1-1+Bst-Exo-3-18 as primers
digest :
Colony Screening
[edit]Date: 2013
DIGEST WITH NdeI , XhoI and BamHI (should get there bands=>350bp, 1500bp,2500bp)
SEND 1144FIX-5 TO SEQUENCE, MISS ONE T and ONE A,NEED TO FIX THEM
cloning
[edit]Date:2013-3-
Fragment 1, use Bst-Exo-1-1 and Bst-Exo-2-8
Fragment 2,use Bst-Exo-2-9 and Bst-Exo-3-18
cut out the right bands, then use Fragment 1and2 as template
Bst-Exo-1-1+Bst-Exo-3-18 as primers
digest :
Colony Screening
[edit]Date: 2013
DIGEST WITH NdeI , XhoI and BamHI (should get there bands=>350bp, 1500bp,2500bp)
Result intepretation/Summary
[edit]Conclusion
[edit]