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Uppsala-Sweden/8 September 2009
From 2009.igem.org
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Since we had quite low concentrations of the ligations we decided to add 20 µl of the ligation mix to the competent cells. The protocoll for the transformation was our standard one. | Since we had quite low concentrations of the ligations we decided to add 20 µl of the ligation mix to the competent cells. The protocoll for the transformation was our standard one. | ||
1) Thaw competent cells on ice (Top 10). | 1) Thaw competent cells on ice (Top 10). | ||
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2) Add ligation mix (20µl). | 2) Add ligation mix (20µl). | ||
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3) Let the cells sit on ice for 30 min. | 3) Let the cells sit on ice for 30 min. | ||
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4) 55 sec heat shock, 42 deg C. | 4) 55 sec heat shock, 42 deg C. | ||
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5) 5 min on ice. | 5) 5 min on ice. | ||
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6) Add 500 µl of SOC | 6) Add 500 µl of SOC | ||
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7) Incubate in shaker at 37 deg C, for 60-70 min. | 7) Incubate in shaker at 37 deg C, for 60-70 min. | ||
Revision as of 16:51, 8 September 2009
Ligation and transformatioin of antisense RNA and other stuff
Performed accordingly: Media:Assembly volume calcuator 090908.xls The concentrations for the purified digestions were quite low but we decided to go for the transformation anyway.
--Anders 16:29, 8 September 2009 (UTC)
Since we had quite low concentrations of the ligations we decided to add 20 µl of the ligation mix to the competent cells. The protocoll for the transformation was our standard one. 1) Thaw competent cells on ice (Top 10).
2) Add ligation mix (20µl).
3) Let the cells sit on ice for 30 min.
4) 55 sec heat shock, 42 deg C.
5) 5 min on ice.
6) Add 500 µl of SOC
7) Incubate in shaker at 37 deg C, for 60-70 min.
