Team:Hawaii/Notebook/2008-08-14

From 2008.igem.org

(Difference between revisions)
(colony pcr)
(marg and grace work added)
Line 7: Line 7:
:* 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 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===
 +
<strong>Margaret</strong>
 +
:*Plasmid preps from [[Team:Hawaii/Notebook/2008-08-14|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===
 +
 +
<strong>Margaret</strong>
 +
 +
:*digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.
== Drylab Work ==
== Drylab Work ==
Line 15: Line 29:
:* Verified sequence of oriT, inserted into pSB1A2. The sequences of oriT and the BioBrick sites are correct.
:* Verified sequence of oriT, inserted into pSB1A2. The sequences of oriT and the BioBrick sites are correct.
 +
===Oligonucleotide Design===
 +
:<strong> Margaret & Grace </strong>
 +
 +
:*We are having trouble with TT, RBS and Promoter ligations, so we decided to synthesize these parts.
 +
:*B0016, I14032, B0030, & B0034
= Discussion =
= Discussion =

Revision as of 03:25, 15 August 2008

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

Things we did today

Wetlab work

Colony PCR

Margaret
  • 5 colonies of rep, P1 lytic region, and aadA(pRL1383a) region, each. pSB1A3 was used as a control.
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 ran another PCR verification on rep, P1, and the omega interposon and afterwards a gel.
File:Rep P1 omega colony pcr.jpg
PCR verification of rep, P1 lytic regbion, and omega interposon ligation from 8-13

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

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


[http://manoa.hawaii.edu/ Sponsor_UHM.gif][http://manoa.hawaii.edu/ovcrge/ Sponsor_OVCRGE.gif][http://www.ctahr.hawaii.edu Sponsor_CTAHR.gif]


Retrieved from "http://2008.igem.org/Team:Hawaii/Notebook/2008-08-14"