Team:Hawaii/Notebook/2008-08-15

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===Made 20X X-gal stock solution (20 mg/ml)===
===Made 20X X-gal stock solution (20 mg/ml)===
:<strong>Grace</strong>
:<strong>Grace</strong>
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===Restriction Digest===
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:<strong>Margaret</strong>
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:*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.
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:*Digested I14032 and the aadA(BB) which was inserted into B0030 so I can ligate these two together.
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===Streak to verify antibiotic resistance===
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:<strong>Margaret</strong>
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:*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 ==
== Drylab Work ==

Latest revision as of 03:16, 16 August 2008

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Things we did today

Wetlab work

Reconstruction of BB-pRL1383a

EtBr stained 1% agarose gel ran at 72V for 1.5 hours. Thirty-five microliters of RE digested BB-pRL1383a and fifty microliters of RE digested J33207 were loaded.
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


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