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Team:Tokyo Metropolitan/Notebook/A04
From 2011.igem.org
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Added 10μl distilled water to wells of J23100 and P0440;distribution kit of iGEM. Then pipetted a couple of times, left it for 5min. After, all collected. | Added 10μl distilled water to wells of J23100 and P0440;distribution kit of iGEM. Then pipetted a couple of times, left it for 5min. After, all collected. | ||
NovaBlue T1R used as copetentcell, not ECOS-competent cell. | NovaBlue T1R used as copetentcell, not ECOS-competent cell. | ||
-Protocol- | -Protocol- | ||
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1. Melt NovaBlue T1R on the ice. | 1. Melt NovaBlue T1R on the ice. | ||
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2. Add 2μl plasumid to NovaBlue T1R, and incubate on the ice for 30 min. | 2. Add 2μl plasumid to NovaBlue T1R, and incubate on the ice for 30 min. | ||
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3. Heatshock at 42℃, 30 sec. | 3. Heatshock at 42℃, 30 sec. | ||
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4. Incubate on the ice for 2 min. | 4. Incubate on the ice for 2 min. | ||
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5. Spread 20-250μl on the plate. | 5. Spread 20-250μl on the plate. | ||
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6. Incubate at 37℃ for 12-14 hours. | 6. Incubate at 37℃ for 12-14 hours. | ||
Revision as of 09:26, 5 October 2011
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-Targeting-
-Methods-
Added 10μl distilled water to wells of J23100 and P0440;distribution kit of iGEM. Then pipetted a couple of times, left it for 5min. After, all collected. NovaBlue T1R used as copetentcell, not ECOS-competent cell.
-Protocol-
1. Melt NovaBlue T1R on the ice.
2. Add 2μl plasumid to NovaBlue T1R, and incubate on the ice for 30 min.
3. Heatshock at 42℃, 30 sec.
4. Incubate on the ice for 2 min.
5. Spread 20-250μl on the plate.
6. Incubate at 37℃ for 12-14 hours.
-Results- E.coli transformed for J23100 and P0440 have been incubating.
-Next experiment- Check the growth of E.coli transformed for J23100 and P0440.(Shimada 1000-) Make LB medium for preculture.(Shimada & Ichikawa 1500-) Preculture E.coli transformed for J23100 and P0440.(Shimada & Ichikawa 2100-)
- Plasmid solution of J23100 and P0440 are keeping in microfuge tubes and in freezer in Cell-biochemistory lab.
Seven plates of LB medium (Amp 50μg/ml)(didn't use) are keeping in cooling room in Biological macinery room.
