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Team:Hawaii/Notebook/2008-08-14
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== Wetlab work == | == Wetlab work == | ||
===Colony PCR=== | ===Colony PCR=== | ||
| + | [[Image: aada_verification_8_14.jpg|right|thumb|300px|PCR verification of aadA from pRL1383a using wrong conditions.]][[Image:rep_P1_omega_colony_pcr.jpg|right|thumb|300px|PCR verification of rep, P1 lytic regbion, and omega interposon ligation from 8-13]] | ||
:<strong> Margaret </strong> | :<strong> Margaret </strong> | ||
:* 5 colonies of rep, P1 lytic region, and aadA(pRL1383a) region, each. pSB1A3 was used as a control. | :* 5 colonies of rep, P1 lytic region, and aadA(pRL1383a) region, each. pSB1A3 was used as a control. | ||
| - | |||
:* I changed the annealing time to 3min 30 seconds instead of the extension time, so the sites were not amplified properly. I ran the aadA(pRL1383a) construct on a gel and got a few bands, none of which were the right size. One of them should have been right though, so I am assuming this ligation did not work. | :* I changed the annealing time to 3min 30 seconds instead of the extension time, so the sites were not amplified properly. I ran the aadA(pRL1383a) construct on a gel and got a few bands, none of which were the right size. One of them should have been right though, so I am assuming this ligation did not work. | ||
:*I ran another PCR verification on rep, P1, and the omega interposon and afterwards a gel. | :*I ran another PCR verification on rep, P1, and the omega interposon and afterwards a gel. | ||
| - | |||
===Culture=== | ===Culture=== | ||
Revision as of 09:06, 15 August 2008
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Colony PCR
File:Rep P1 omega colony pcr.jpg
PCR verification of rep, P1 lytic regbion, and omega interposon ligation from 8-13
- Margaret
- 5 colonies of rep, P1 lytic region, and aadA(pRL1383a) region, each. pSB1A3 was used as a control.
- I changed the annealing time to 3min 30 seconds instead of the extension time, so the sites were not amplified properly. I ran the aadA(pRL1383a) construct on a gel and got a few bands, none of which were the right size. One of them should have been right though, so I am assuming this ligation did not work.
- I ran another PCR verification on rep, P1, and the omega interposon and afterwards a gel.
Culture
Margaret
- Plasmid preps from yesterday did not yield very high quantities. I am setting up the culture for oriV1-4 and aadA(BB) 4 so I can do another plasmid prep tomorrow.
Restriction Digest
Margaret
- digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.
Drylab Work
Sequencing
- Margaret
- Verified sequence of oriT, inserted into pSB1A2. The sequences of oriT and the BioBrick sites are correct.
Oligonucleotide Design
- Margaret & Grace
- We are having trouble with TT, RBS and Promoter ligations, so we decided to synthesize these parts.
- B0016, I14032, B0030, & B0034
Discussion
Quote of the Day
happy chickens taste better - KGB
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
