Team:Hawaii/Notebook/2008-08-14

From 2008.igem.org

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(Culture)
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===Restriction Digest===
===Restriction Digest===
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:<strong>Margaret</strong>
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<strong>Margaret</strong>
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:*digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.
:*digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.
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:<strong>Grace</strong>
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:* Checked Krystle's RE digests from last night on a gel
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:* RE digested:
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::* J33207, J04430, BB-pRL1383a with EcoRI and PstI
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::* pRL1383a, J33207 with HindIII and BamHI (Roche, from SC)
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::* BB-pRL1383a with HindIII and BamHI (from lab -20C)
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:* Ran RE digests on a gel to purify segments of interestt
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::* Also ran RE digested B0015 from Krystle
== Drylab Work ==
== Drylab Work ==

Revision as of 09:09, 15 August 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

Colony PCR

PCR verification of aadA from pRL1383a using wrong conditions.
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.
Grace
  • Checked Krystle's RE digests from last night on a gel
  • RE digested:
  • J33207, J04430, BB-pRL1383a with EcoRI and PstI
  • pRL1383a, J33207 with HindIII and BamHI (Roche, from SC)
  • BB-pRL1383a with HindIII and BamHI (from lab -20C)
  • Ran RE digests on a gel to purify segments of interestt
  • Also ran RE digested B0015 from Krystle

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


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