IGEM:IMPERIAL/2006/LabCalendar/2006-8-18

Maxipreps

 * 12D - for J37020 ligation
 * 5M - for ligating into aiiA construct when primers arrive
 * J37016 - so a sample can be sent for sequencing

Sequencing

 * J37016 sent for sequencing
 * 1 DNA
 * 12.5 pmol I13504 primer (CAA CTA GCA GAC CAT TAT C)

Minipreps

 * J37015RS
 * We have innoculated a 100 mL culture in the fridge to be maxipreped on Monday
 * All cultures were successful; however, we took only colony 4 since it showed the best looking bands on the gel (yes, I guess we do have to be discriminatory)
 * 1M
 * Band at around 2000 showing the plasmid, the part itself is too small and may have run too far, so we assume that it is correct
 * This part can be maxipreped on Monday along with J37015RS
 * 4G
 * Successful miniprep, we can start testing this on Monday as it is an entire part ready to be testing. Please ensure that you culture in Kanamycin LB (located in fridge)
 * 2H
 * Unsuccessful, needs redoing
 * Second miniprep unsuccessful. Have to let MIT know we have been struggling to ligate part 2H and that we suspect that it may involve a mutation in the Spe1 site.



Ligations

 * J37031 = J37019 + B0015 (1I)
 * J37032 = R0062 (9G) + I13504 (12D)
 * 12D on the gel after digestion was not clean, this might pose a small problem, but I cut out the smaller band anyway that look around 800 bp (compared to the J37019 which is also about 800 bp).
 * Electroporations this afternoon