Revision as of 09:09, 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

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

happy chickens taste better - KGB

@@ Line 16: / Line 16: @@
 ===Restriction Digest===
+:<strong>Margaret</strong>
-<strong>Margaret</strong>
 :*digest of I14032 with SpeI and PstI. I want to use this to ligate to the aadA(BB) construct.
+:<strong>Grace</strong>
+:* Checked Krystle's RE digests from last night on a gel
+:* RE digested:
+::* J33207, J04430, BB-pRL1383a with EcoRI and PstI
+::* pRL1383a, J33207 with HindIII and BamHI (Roche, from SC)
+::* BB-pRL1383a with HindIII and BamHI (from lab -20C)
+:* Ran RE digests on a gel to purify segments of interestt
+::* Also ran RE digested B0015 from Krystle
 == Drylab Work ==