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Team:Hawaii/Notebook/2008-08-27
From 2008.igem.org
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Things we did today
Wetlab work
Construction of p+r
- Grace
- Ran RE digested products on gel
- nir had three bands. Band at ~730bp = ?
- BB-pRL1383a -- still couldn't see cut out of GFP. Assumed double digest successful, too little DNA to see GFP.
- J33207 wrong size
- Extracted nir, C0012 vector, BB-pRL1383a from gel
- 3A assembly (ligation) of C0012 vector (pSB1A2) with:
- nir and rbs (B0034)
- plac and rbs(B0034)
- pSB1A2 (negative control)
- Transformed DB3.1 cells using 5 μl ligation reaction
Transformation
- Margaret
- ligation products from yesterday were transformed into DH5-a cells, pSMC121 was used as a positive control
- rep+pSB1A3 (1:1)
- rep+pSB1A3 (1:3)
- rep+pSB1A3 (1:6)
- P1 lytic +pSB1A3 (1:1)
- P1 lytic +pSB1A3 (1:3)
- P1 lytic +pSB1A3 (1:6)
- [Plac B0030]+pSB1A3 (1:3)
- [Plac B0030]+pSB1A3 (1:6)
- [Plac B0034]+pSB1A3 (1:6)**ran out of vector
Sequencing
- Margaret
- in preparation for sequencing, a PCR was run on the following plasmids:
oriV1-4, aadA(BB)5, aadA(BB)9, aadA(pRL)3, aadA(pRL)6, omega interposon
- Note: In lane five, it looks like I added oriV instead of aada(BB)5. In the omega lane, there is a ladder. I will still sequence these.
- Grace
- Prepared nir+rbs and rbs+GFP constructs for sequencing
Discussion
Quote of the Day
History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson
