Team:Hawaii/Notebook/2008-08-15

From 2008.igem.org

(Difference between revisions)
(→Reconstruction of BB-pRL1383a)
(margarets list)
 
(3 intermediate revisions not shown)
Line 4: Line 4:
== Wetlab work ==
== Wetlab work ==
===Reconstruction of BB-pRL1383a===
===Reconstruction of BB-pRL1383a===
-
[[Image:081508REdigest.jpg|right|thumb|200px|EtBr stained 1% agarose gel ran at 72V for ... Thirty-five microliters of RE digested BB-pRL1383a and fifty microliters of RE digested J33207 were loaded.]]
+
[[Image:081508REdigest.jpg|right|thumb|200px|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.]]
:<strong> Grace</strong>
:<strong> Grace</strong>
Line 13: Line 13:
::* Ladder did not run well. Bands not resolving well.
::* Ladder did not run well. Bands not resolving well.
:* Redid PCR and RE digests with XbaI and PstI
:* Redid PCR and RE digests with XbaI and PstI
 +
 +
===Made 20X X-gal stock solution (20 mg/ml)===
 +
:<strong>Grace</strong>
 +
 +
 +
===Restriction Digest===
 +
:<strong>Margaret</strong>
 +
 +
:*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===
 +
:<strong>Margaret</strong>
 +
 +
:*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 ==
Line 24: Line 38:
= Quote of the Day =
= Quote of the Day =
-
<blockquote>''History is the only laboratory we have in which to test the consequences of thought.'' - Étienne Gilson</blockquote>
+
<blockquote>''I'm waiting for the terminator to come in...'' - Krystle</blockquote>
{{Team:Hawaii/Footer}}
{{Team:Hawaii/Footer}}
__NOTOC__
__NOTOC__

Latest revision as of 03:16, 16 August 2008

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

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


[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/2008-08-15"