Team:Hawaii/Notebook/2008-09- 6

= Things we did today =

Construction of B_H_R Vector

 * Margaret


 * Yesterday's Restriction Digest
 * Plac+B0030 and Plac+B0034: ethanol precipitation []
 * pSB1A3, pSB3K3, J33207: de-phosphorylated using SAP


 * Gel Quantification


 * Some of these products did not appear on the gel, so other methods will be used to generate them.


 * Plac+B0030 and Plac+B0034 did not show up on the gel, so the ligation is being performed tonight, pSB1A3 and pSB3K3 plasmid preps from 9/4 showed a good band.
 * Ligation 1:1, followed by Restriction digest with E,S,& P (to cut up any polymers formed and to leave E & S overhang):
 * Plac + B0030 (500ng)
 * Plac + B0030 (100ng)
 * Plac + B0034 (500ng)
 * Plac + B0030 (100ng)
 * pSB3K3 restriction digested with E, P

OriT test

 * Margaret
 * The biobrick form of OriTr did not show up on the above mentioned gel, so I am:
 * PCR amplifying it from the biobrick paper
 * Extracting DNA from the biobrick registry (1004, 10D) and transforming into DH5-alpha cells. The negative control is pSB1A3-dephosphorylated, the positive control is pSB1A3, from a plasmid prep.

Culturing

 * Margaret
 * DH5-alpha (for making competent cells)
 * rep, J33207, pSB3K3, and omega interposon

= Discussion =

= Quote of the Day = "History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson"