DNA Precipitation

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Procedure)
(Notes)
Line 20: Line 20:
==Notes==
==Notes==
-
#List troubleshooting tips here. 
+
<font face="courier"><nowiki>'''*~~~~''':</nowiki></font>
-
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
+
-20&deg;C for an hour is fine for using larger (1mL of bacterial culture, plasmid) amounts of DNA
-
#Anecdotal observations that might be of use to others can also be posted here. 
+
-
Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
+
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.
==References==
==References==

Revision as of 17:18, 22 April 2008

Contents

Overview

Replace this sentence with a brief description of the protocol and its goal.

Materials

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)

Procedure

  1. 1/10 volume NaOAc.
  2. 1ul Glycogen.
  3. 2 volumes EtOH.
  4. -20°C overnight or 30min -80°C.
  5. Centrifuge >12k G for at least 15 mins.
  6. Remove supernatant
  7. Resuspend in desired volume of water/buffer

Notes

'''*~~~~''': -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.

References

Relevant papers and books

  1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Jacob-JMB-1961]
  3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Ptashne-Genetic-Switch]
All Medline abstracts: PubMed HubMed

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


Personal tools