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Protocol for DNA Purification after Enzymatic Reaction by Phenol/Chloroform Extraction and Alcohol Precipitation

1. Mix your sample with 0.5 volume of TE-saturated phenol and 0.5 volume of chloroform. Then, centrifuge (10,000 rpm, 5 min, room temperature).
2. Transfer the upper phase to a fresh tube. Add an equal volume of chloroform and mix. Then, centrifuge (10,000 rpm, 5 min, room temperature).
3. Transfer the upper phase to a fresh tube. Add 1/10 volume of 3 M sodium acetate or 2 M sodium chloride.
4. Add an equal volume of isopropanol or 2.5 volumes of ethanol to precipitate DNA.
5. Incubate the mixture for 30-60 min at -20°C.
6. Centrifuge for 10 min at 10,000 rpm. Then discard the supernatant and rinse the pellet twice with 70% cold ethanol.
7. Air-dry the pellet. Dissolve in water, nuclease-free or TE buffer for further use.

Note
Use Glycogen to maximize the yield of DNA during precipitation, see detailed protocol.

Ordering Information

RNase A/T1 Mix Proteinase K GeneJET™ Plasmid Miniprep Kit Genomic DNA Purification Kit Glycogen Water, nuclease-free

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Updated gegužės 09, 2007 17:26