Revision as of 09:14, 15 August 2008

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Things we did today

PCR verification of aadA from pRL1383a using wrong conditions.

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.

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.

Margaret

digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.

Grace

EtBr stained 2% agarose gel ran at 72V for 1.5 hours. Thirty microliters of the RE digests were loade dinto each well.

Margaret

Verified sequence of oriT, inserted into pSB1A2. The sequences of oriT and the BioBrick sites are correct.

Margaret & Grace

We are having trouble with TT, RBS and Promoter ligations, so we decided to synthesize these parts.
B0016, I14032, B0030, & B0034

Gel box 1 is the only one that works properly. The unlabeled gel box and box #2 may cause gels to run excessively slow, bands to blur/smear, or other weird things. Amps is really low on these boxes. All covers/power sources are good.

happy chickens taste better - KGB

[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]

@@ Line 21: / Line 21: @@
 :<strong>Grace</strong>
-[[Image:081408REdigests.jpg|right|thumb|200px|EtBr stained 2% agarose gel ran at 72V for 1.5 hours. Thirty microliters of the RE digests were loade dinto each well.]]
 :* Checked Krystle's RE digests from last night on a gel
 :* RE digested:
@@ Line 29: / Line 29: @@
 :* Ran RE digests on a gel to purify segments of interest
 ::* Also ran RE digested B0015 from Krystle
+[[Image:081408REdigests.jpg|right|thumb|200px|EtBr stained 2% agarose gel ran at 72V for 1.5 hours. Thirty microliters of the RE digests were loade dinto each well.]]
 == Drylab Work ==