Team:Heidelberg/Templates/DelH week26
From 2013.igem.org
Contents |
21-10-2013
Preparation of large plasmids
6 ml of clones A1, A4, A7, B4
Restriction Digest of Preps with PvuI
0.5 µl PvuI-HF, 2 µl CutSmart, 1 µl 1:10 Prep (1-2 µg)
2 hours of incubation
expected bands: 11.6 kbp, 5.8 kbp, 4.6 kbp
--> only clone A1 was positive
--> A1 was send to sequencing with primer DN07
Screening colonies of pSB4K5+DelH (without promotor)
46 colonies
0.5 Primer (FS73 and FS95)
5 µl Dream-Taq
4 µl ddH2O
Test if electrocompetent BL21-DE3 with DelRest and pIK8.6 contain DelRest
Screening of plasmid pFSN-D8w (positive control), Triple clone and electrocompetent BL21-DE3 with DelRest and pIK8.6
0.5 µl FS_14
0.5 µl VR
4 µl ddH2O
5 µl Dream Taq
Results:
- All three screenings were positive. Consequently our electrocompetent BL21-DE3 with DelRest and pIK8.6 really contain DelRest.