Team:Hawaii/Notebook/2008-08-15

= Things we did today =

Reconstruction of BB-pRL1383a

 * 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).

Editing team website

 * Grace

= Discussion =
 * DH5&alpha; 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"