The goal of this protocol is to precipitate DNA, as the name says. This is usually coupled with phenol chloroform extraction and is used as a way of purifying nucleic acids. This can also be used as a method for changing what solution or buffer your nucleic acid is in.
This protocol also works for RNA precipitation (take care to us RNAse free materials in this case).
List reagents, supplies and equipment necessary to perform the protocol here. For those materials which have their own OWW pages, link to that page. Alternatively, links to the suppliers' page on that material are also appropriate.
- 3M NaOAc pH 5.2
- EtOH 95%
- Glycogen (optional)
- 1/10 volume NaOAc.
- 1ul Glycogen.
- 2 volumes EtOH.
- -20°C overnight or 30min -80°C.
- Centrifuge >12k G for at least 15 mins.
- Remove supernatant
- Resuspend in desired volume of water/buffer
- Ryan R Hurtado 17:21, 22 April 2008 (EDT):
-20°C for an hour is fine for using larger (1mL of bacterial culture, plasmid) amounts of DNA
The DNA pellet will not always be visible depending on how much DNA you are precipitating. So always take care in loading your samples in the centrifuge to remember the direction they are facing. The DNA pellet will be on the part of the tube facing the outside of the centrifuge.
This protocol will precipitate all nucleic acids, not just DNA. If you do not want RNA in your sample, one of the many ways to deal with it is to simply resuspend in TE + RNAse at the last step and leave it at room temperature for 15mins-1hr.
Relevant papers and books
- Goldbeter A and Koshland DE Jr. . pmid:6947258.
- JACOB F and MONOD J. . pmid:13718526.
- Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164.
- Who has experience with this protocol?
or instead, discuss this protocol.