1265 pET30b-Tks
Cloning of 1265 pET30b-Tks
[edit]Purpose: Cloning of 1265 pET30b-Tks
Date: July 13, 2012
Notebook:
Page:
Clone ID: 1265
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: July 13, 2012
Gene synthesis of 4 fragments and then join by PCR, expected size 2.4 kb
Not correct size, need to reamplify on Monday
Procedure: 4 PCR reactions for each fragment, (1 uL 10 uM mix, 1 uL for template) with PHUSION enzyme, 10 cycles. Then took 6 uL from each fragment, added 1 uL more of PHUSION enzyme and 1 uL of Tks-1-1 and Tks3-18 (primers), PCR for 20 more cycles with 2:30 minutes extension time
Looks like fragments did not join together? Band is around ~600 bp, which is the size of each fragment...
Colony Screening
[edit]Result intepretation/Summary
[edit]Conclusion
[edit]==
Cloning of 1265-pET30b-Tks
[edit]Purpose: Cloning of 1025-pET28b-SM2
Date: 2012-7-31---2012-8-2
Notebook:0
Page:
Clone ID:1265
Experimenters: yingying
Results
[edit]cloning
[edit]
Gene synthesis of 4 fragments
2nd PCR join four fragement together by PCR method
Colony Screening
[edit]Date: 2012-8-2
Make miniprep and digest with
no insert