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Uppsala-Sweden/8 September 2009
From 2009.igem.org
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--[[User:Anders|Anders]] 16:29, 8 September 2009 (UTC) | --[[User:Anders|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. | + | 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. |
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1) Thaw competent cells on ice (Top 10). | 1) Thaw competent cells on ice (Top 10). | ||
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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. | ||
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| + | 8) Plate and grow over night. | ||
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| + | --[[User:Flormane|Flormane]] 07:56, 9 September 2009 (UTC) | ||
Latest revision as of 07:56, 9 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.
8) Plate and grow over night.
--Flormane 07:56, 9 September 2009 (UTC)
