Team:Chiba/Calendar-Home/1 September 2008

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31 August 2008 <|> 2 September 2008

Team:Input
Gel Check
 * Ptet＋RBS＋cI

Transformation(Double Transformation)
 * -Ptet-cI-pMB1-Amp-,-PcI-GFP-p15A-Cm-(JW1908Δflic)

Digestion test
 * -Ptet-cI-pMB1-Amp-

UV irradiation test
 * two plasmids from (JW1908)
 * -Ptrc-LuxR-Plux-cI-colE1-Amp-
 * -PcI-GFP-p15a-Cm-

OD:UV⊕:5.21,UV⊖(negative control):5.38
 * 1) Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
 * 1) moved the cultures to small plates and started UV-irradiation.(wavelength:254nm,distance from the UV lamp to the cultures were 7.5cm.Put the negartive control in a dark place.
 * 2) covered the plates with polyethylene wrap.
 * 3) after irradiation(Sample:0min,10min,30min,1h,2h,4h,6h,8h)(Negative Control:0h,1h,4h,8h),we agitate the culture and took 20&mu;l from each other
 * 4) diluted each cultures with LB-ampicillin medium 104-fold and 105-fold (volume/volume).
 * 5) incoculated 20μl of the cultures and incubated for 12 hours at 37 degrees.
 * 6) counted the CFU(determined viable cell count).

Viable cell count

Team:Communication

 * (31/8)--->Gel Check


 * --->Gel extract


 * --->zymo
 * insert-1(BBa_I9026) -> 7μL
 * insert-2(BBa_I9030) -> 7μL
 * insert-3(BBa_S03154) -> 7μL
 * vector-4(BBa_R0010) -> 15μL


 * --->SAP
 * vector-4(BBa_R0010)


 * --->Zymo
 * vector-4(BBa_R0010) -> 20μL


 * --->Gel Check


 * --->Ligation


 * --->Transformation
 * Competent cells : XL10GOLD 30μL
 * Transformed the following and grew on new ampicillin plates.
 * BBa_K084009(Plac+RBS+RhlI+LVA, Amp) -> 628 colonies
 * BBa_K084010(Plac+RBS+CinI+LVA, Amp) -> 500 colonies
 * insert-1(RBS+RhlI+LVA) -> 9 colonies
 * insert-2(RBS+CinI+LVA) -> No colonies on the plate
 * vector-4(Plac, Amp) -> 186 colonies


 * --->(2/9) Colony PCR

Transformation
 * Competent cells : JW1908 40μL
 * Transformed the following and grew on new ampicillin plates.
 * BBa_K084007(Plac+RBS+LasI)
 * BBa_K084008(Plac+RBS+RhlI)
 * BBa_T9002(Ptet+RBS+LuxR+GFP)


 * --->(2/9)Liquid Culture

Team:Output
Ligation
 * vector:BBa_R0010 insert:BBa_J52008(1)
 * negative control:BBa_R0010(2)

-->R/T 2hour

Transformation


 * BBa_R0079
 * BBa_R0071
 * BBa_R0077
 * BBa_R0078
 * BBa_R0062