IGEM:Peking/2007/Switch: DNA rescue by precipitation

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(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...)
Current revision (11:02, 22 October 2007) (view source)
(DNA沉淀回收)
 
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==DNA沉淀回收==
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==DNA rescue by precipitation==
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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。片断越短需要沉淀时间越长。<100bp的片断则应用等体积的异丙醇+1/10体积的3M NaAC沉淀过夜。
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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.
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2. 12,000rpm离心15min。
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2. centrifuge for 12,000rpm, 15min
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3. 弃去上清。
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3. Discard the supernatant
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4. 加入200uL 70%乙醇清洗管壁。
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4. Add 200uL 70% enthol to wash.
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5. 12,000rpm离心10min。
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5. centrifuge for 12,000rpm, 10min
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6. 弃上清。
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6. Discard the supernatant
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7. 在37℃烘干。
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7. Dry it under 37 degrees
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8. 溶于适当体积(20uL)的ddH2O中。
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8. Dissovle it in some volume of ddH2O, say 20uL.

Current revision

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.

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