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Team:Hawaii/Notebook/2008-08-15
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
- Grace
- PCR amplification of J33207
- ExoSAP'd PCR product
- RE digested of J33207 and BB-pRL1383a (w/ GFP) with EcoRI and PstI
- Ran digests on a 1% agarose gel at 72V for 1.5 hours.
- Ladder did not run well. Bands not resolving well.
- Redid PCR and RE digests with XbaI and PstI
Made 20X X-gal stock solution (20 mg/ml)
- Grace
Restriction Digest
- Margaret
- Digested J33207 (contains lacZ for blue/white screening), oriT--> these will be ligated so i can test the origin of transfer in a conjugation experiment.
- Digested I14032 and the aadA(BB) which was inserted into B0030 so I can ligate these two together.
Streak to verify antibiotic resistance
- Margaret
- On an LB plate containing Sm&Sp, the omega ligation (from 8-12). This will be a verification that the omega ligation worked. As a positive control I streaked pSMC121 which contains the omega interposon, and as a negative control I streaked pSB1A2 (in case the plates have lost their effectiveness).
Drylab Work
Editing team website
- Grace
Discussion
- DH5α does not carry lacIq; therefore, IPTG is not neccessary to induce the lac promoter.
- IPTG is necessary for induction in DB3.1 cells.
Quote of the Day
I'm waiting for the terminator to come in... - Krystle
