1217 pDB.His.PfuMBP-(3C): Difference between revisions
No edit summary |
(No difference)
|
Latest revision as of 09:15, 21 August 2012
Cloning of 1217 pDB.His.PfuMBP-(3C)
[edit]Purpose: Cloning of 1217 pDB.His.PfuMBP-(3C)
Date: June 26, 2012
Notebook:
Page:
Clone ID: 1217
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]
Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1), looks like it is not digested. Ran out of 1129 vector, growing new from gycerol stock. Will redo tomorrow. Stored annealed oligos in box.
Date: June 27, 2012
Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1) again, same 2 bands happened.
Digested vector separately with BamHI and Nde1, digest with NEB enzymes
Digested with NEB BamHI and Nde1
Date: August 17, 2012
Digested pDB.His.PfuMBP 1129 with Nde1 + BamH1 (see lane 1)
Colony Screening
[edit]
Digested with RE Pst + Xba1
Very light band at expected 1383 bp, very unclear
Sequencing result is not good, need to do it again.
Date: August 20, 2012
Digest with Pst + Xba1--->get a band of 1272 bp
Sent sample 5 to sequencing
Clones 1, 2, 3, 5 may be correct
Result intepretation/Summary
[edit]
Conclusion
[edit]