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Team:Hawaii/Notebook/2008-08-16
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Reconstruction of BB-pRL1383a (cont.)
- Grace
- Ran digests on a 1% agarose gel
- J33207 bands @ ~900bp and smudge ~100bp. ~100bp=VF/VR? Too much DNA loaded = band ran slower?
- BB-pRL1383a band @ ~10kb. No band ~750bp. XbaI/PstI did not cut??
- Extracted bands from gel
- Ligated:
- 1 μl XbaI/PstI digested BB-pRL1383a with 3 μl XbaI/PstI digested J33207
- 1 μl XbaI/PstI digested BB-pRL1383a to itself (negative control)
- Transformed into DH5α
- Used 5 μl ligation reaction to transform
- Plated on LB+sp100+X-gal and LB+sp100+sm50+IPTG+X-gal
- To verify if IPTG is needed for lacZ expression
Plasmid preps
- Grace
- Resuspended plasmid preps in 50 μl TE buffer and stored in -20C freezer (long green tray)
Discussion
Quote of the Day
She grew on him like she was a colony of E. coli, and he was room temperature Canadian beef. - Courtesy of Help.com
