IGEM:Peking/2007/Switch: DNA rescue by precipitation: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Chen Chongyi (talk | contribs) (New page: ==DNA沉淀回收== 1． 向体系中加入2倍体积的100%乙醇，1/10体积的3M NaAC，放置适当时间可使DNA沉淀。对于载体，一般不短于30min。片断越短需要沉...) |
Chen Chongyi (talk | contribs) |
||
| Line 1: | Line 1: | ||
== | ==DNA rescue by precipitation== | ||
1． | 1． Add 2 volume times of 100% ethanol in the system, 1/10 volume times of 3M NaAc. Some time later DNA will precipitate. For vectors, generally speaking the time is shorter than 30 minutes. For fragments shorter, the time needed for precipitation is longer. If some fragment shorter than 100bp need precipitate, same volume of isopropanol and 1/10 volume of 3M NaAc should be added and precipitation process need overnight. | ||
2． | 2． centrifuge for 12，000rpm, 15min | ||
3． | 3． Discard the supernatant | ||
4． | 4． Add 200uL 70% enthol to wash. | ||
5． | 5． centrifuge for 12，000rpm, 10min | ||
6． | 6． Discard the supernatant | ||
7． | 7． Dry it under 37 degrees | ||
8． | 8． Dissovle it in some volume of ddH2O, say 20uL. | ||
Latest revision as of 08:02, 22 October 2007
DNA rescue by precipitation
1． Add 2 volume times of 100% ethanol in the system, 1/10 volume times of 3M NaAc. Some time later DNA will precipitate. For vectors, generally speaking the time is shorter than 30 minutes. For fragments shorter, the time needed for precipitation is longer. If some fragment shorter than 100bp need precipitate, same volume of isopropanol and 1/10 volume of 3M NaAc should be added and precipitation process need overnight.
2． centrifuge for 12，000rpm, 15min
3． Discard the supernatant
4． Add 200uL 70% enthol to wash.
5． centrifuge for 12，000rpm, 10min
6． Discard the supernatant
7． Dry it under 37 degrees
8． Dissovle it in some volume of ddH2O, say 20uL.
