Team:Hawaii/Notebook/2008-08-14

From 2008.igem.org

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(Quote of the Day)
(Colony PCR)
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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.
-
[[Image: aada_verification_8_14.jpg|right|thumb|300px|PCR verification of aadA from pRL1383a using wrong conditions.]]
 
:* 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.
-
[[Image:rep_P1_omega_colony_pcr.jpg|right|thumb|300px|PCR verification of rep, P1 lytic regbion, and omega interposon ligation from 8-13]]
 
===Culture===
===Culture===

Revision as of 09:06, 15 August 2008

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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.

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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