Team:KIT-Kyoto/Notebook-week3
From 2012.igem.org
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![](https://static.igem.org/mediawiki/2012/0/02/KIT-KyotoB.jpg)
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August 20th*TNFA and API2 1. Reproduction of DNA from gel We did agarose gel electrophoresis (0.7% gel). Composition
We separated 54uL×2 from 108uL (sample) and we did electrophoresis (54uL sample and 1kb marker 5uL).We separated gel containing DNA. Results ![]() We isolated DNA from these gels by QIA quick Gel Extraction Kit. Finally we melt DNA to TE Buffer(40uL) August 21st*TNFA and API2 1. Density measurement of attB TNFAIP3 We did agarose gel electrophoresis (0.7% gel). (1kb marker-3uL, 5times dilution attB TNFAIP3-1uL (make August 20 ), 10times dilution-1uL, 1kb marker-2uL, 15times dilution-1uL, 20times dilution-1uL, 1kb marker-1uL ) Results ![]() We estimated attB TNFAIP3(we make this time) is 35ng/uL 2. BP reaction We adjusted solution (vials) on 1.5mL tube to next composition.
We added BP Clonase Ⅱ enzyme mix-2uL to this vials. We incubated these vials for 2hour at 25˚C. Next we did BP reaction 3. Transformation We added BP TNFAIP3-2uL to XL1-Blue-100uL and we did Transformation. Finally we spread 10uL, 40uL and 100uL on plate of LB Kanamycin(+) and we keep 37℃ overnight. August 22nd*TNFA and API2 We separated six colonies from 40uL spread plate of BP TNFAIP3 (make August 21 ) . We cultivated these colonies on 2.5mL Kanamycin(+) LB culture medium and we kept shaking this medium overnight. August 23rd*TNFA and API2 ![]() *Parts August 24th*TNFA and API2 *Parts August 25th*TNFA and API2 *Parts
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August 26th*TNFA and API2 The female-virgin flies (w; Δ2,3) and male flies (yw) were collected for microinjection. The collected flies were kept in the 25℃ incubator separately. *Parts ![]() |