Team:UNIPV-Pavia/Calendar/August

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Revision as of 18:59, 9 August 2010

AUGUST

Bio-Lab activity proceeds!

Select the day you are interested in to find out the details!

DATE	ACTIVITY
August, 1st	44° Bio-Lab - I20 and I21 plates showed colonies. Some were picked, inoculated and grown in proper medium.
AUGUST: WEEK 1
August, 2nd	45° Bio-Lab - I20-1/2/3 and I21-1/2/3 ligations sent sequencing. <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> resuspended from iGEM 2010 Distribution Kit. Trasformation of pAH123 and <partinfo>BBa_J72008</partinfo> helper into MC1061 and MG1655.
August, 3rd	46° Bio-Lab - <partinfo>BBa_R0062</partinfo> and <partinfo>BBa_K081009</partinfo> single colony pick from plates. MC1061 and MG1655 colonies transformed with helpers were picked from plates and inoculated and and incubated in proper medium. PCR from colony (MG1655 and MC1061) using synthesized primers to check them and to verify a negative control for integration.
August, 4th	47° Bio-Lab - Glycerol stocks for MC1061 and MG1655 strains transformed with pAH123 or <partinfo>BBa_J72008</partinfo> helper plasmids. Miniprep of MC1061 and MG1655 to check again the presence of helper plasmids. Digestion with SpeI and gel run. We got positive samples but a bad run. So we picked again from plates, this time single colonies.
August, 5th	48° Bio-Lab - Screening for MG1655 and MC1061 carrying <partinfo>BBa_J72008</partinfo> or pAH123 helpers: miniprep of new picked colonies MC1061 and MG1655, digestion with SpeI and gel run. New ligation cycle for phasins (I20-new, I21-new).
August, 6th	49° Bio-Lab - Transformation of I20-new and I21-new ligations into E. coli Dh5-alpha. Competentization of MG008/123 and MC008/123 strains selected previous day.
August, 7th	50° Bio-Lab -
August, 8th	51° Bio-Lab -
AUGUST: WEEK 2
August, 9th	52° Bio-Lab - Phasins plates showed very few colonies (17 for I20-new and 12 for I21-new): they were all picked and let grow in LB+Amp 100ug/ml. In the evening we made glycerol stocks and re-filled the tubes for the screening of the following day. Not to loose time we performed again PCR-amplification/modification of <partinfo>BBa_K208001</partinfo>. We checked LB+Cm 6 ug/ml agar plates with MC1061, MG1655, MC008, MG008, MC123, MG123 (none of them should grow). Check for transformation efficiency of MC123/008 and MG123/008.
August, 10th	53° Bio-Lab -
August, 11th	54° Bio-Lab -
August, 12th	55° Bio-Lab -
August, 13th	56° Bio-Lab -
August, 14th	57° Bio-Lab -
August, 15th	58° Bio-Lab -

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 We checked LB+Cm 6 ug/ml agar plates with MC1061, MG1655, MC008, MG008, MC123, MG123 (none of them should grow).
+Check for transformation efficiency of MC123/008 and MG123/008.
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