IGEM:IMPERIAL/2006/LabCalendar/2006-9-5

 TO DO: *Testing of J37016, S01656 *Finish ligation for J37022 & electroporate (has it been finished yesterday?) *Finish ligation for Dr. Frays aiiA into the T hanging plasmid & electroporate *Culture J37036 & J37018 from plate in incubator for maxi & mini on Wednesday *Culture T9002 from plate in incubator for new testing & making frozen stocks

Western Blot

 * There was no discernable difference in the proteins produced by the induced and non-induced cells
 * This indicates that there is something wrong with the mechanism involving S01656 expression, possibly its promoter
 * This does not however mean that the aiiA is wrong necessarily - testing of J37022 should clarify this.



Testing of S01656

 * Was started with the revised protocol but after results from Western Blot came out, was abandonded
 * Is not going to be tested any more since no protein seems to be transcribed at all

AiiA Ligation/electroporation

 * Samples run on gel and adequate bands cut out.
 * Glass milk purified and ligated into T9 overhanging vector
 * Forms New J37023
 * Electroporated in the afternoon

LoxP PCR

 * The small PCR fragment is not being amplified

Testing S01656 & J37016

 * Testing was not conducted as western blot indicates that there may be something wrong with the S01656 expression mechanism.
 * J37016 was being cultured for the S01656 testing. This test, too, was not finished since we are no longer carrying out testing of S01656. We have 3 good sets of data for J37016 so far.

Culturing

 * Cultured up 4 colonies of J37036 in 110ml of LB Amp for maxi and mini
 * Cultured up 2 colonies of J37018 in 110ml of LB Amp for maxi and mini
 * Cultured up 2 colonies of T9002 in 2ml of LB Amp for tesing?
 * Cultured up J37015 in 2ml of LB Amp for testing
 * Cultured up J37016 in 2ml of LB Amp for testing