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Team:Heidelberg/Templates/DelH week26
From 2013.igem.org
(Difference between revisions)
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==Restriction Digest of Preps with PvuI== | ==Restriction Digest of Preps with PvuI== | ||
0.5 µl PvuI-HF, 2 µl CutSmart, 1 µl 1:10 Prep (1-2 µg) <br/> | 0.5 µl PvuI-HF, 2 µl CutSmart, 1 µl 1:10 Prep (1-2 µg) <br/> | ||
| - | 2 hours of incubation | + | 2 hours of incubation <br/> |
| + | expected bands: 11.6 kbp, 5.8 kbp, 4.6 kbp <br/> | ||
| + | --> only clone A1 was positive <br/> | ||
| + | --> A1 was send to sequencing with primer DN07 | ||
==Screening colonies of pSB4K5+DelH (without promotor)== | ==Screening colonies of pSB4K5+DelH (without promotor)== | ||
46 colonies <br/> | 46 colonies <br/> | ||
0.5 Primer (FS73 and FS95) <br/> | 0.5 Primer (FS73 and FS95) <br/> | ||
| + | 5 µl Dream-Taq <br/> | ||
| + | 4 µl ddH<sub>2</sub>O | ||
| + | ==Test if electrocompetent BL21-DE3 with DelRest and pIK8.6 contain DelRest== | ||
Revision as of 15:33, 21 October 2013
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
