Team:Hawaii/Notebook/2008-08-27

= Things we did today =

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 &mu;l 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"