Mouse Heart Tissue Harvest for DNA Microarrays: Difference between revisions

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==Overview==
==Overview==


Replace this sentence with a brief description of the protocol and its goal.
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==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.
 
*supply 1 (i.e. tubes of a certain size? spreaders?)
*reagent 1
*X μL reagent 2
**component A (reagent 2 is made up of multiple components)
**component B
*equipment 1
*equipment 2


==Procedure==
==Procedure==

Revision as of 12:03, 16 June 2009

Overview

List of Reagents not yet complete

Procedure

1. Anesthesize mouse with 0.02 ml 2.5% Avertin /gram body weight. 2. Make incisions through the skin and the peritoneum and cut out approximately 100-200 mg of the tissues to be analyzed. 3. Immediately rinse the tissue in 1x PBS, blot dry, and weigh. 4. Transfer the tissue into an eppendorf tube and freeze in liquid nitrogen. 5. After all the samples are collected, they may be stored at -70°C or used for immunoprecipitation.

2.5% Avertin (Chem Name = Tribromoethanol, Fluka #90710) 

   Dilute 100% Avertin in warm PBS (55°C or higher) Filter sterilize, wrap in foil, store frozen aliqouts at -20°C Aliqouts good for ~3 weeks at 4°C

100% Avertin 

   Dissolve Avertin in tert-amyl alcohol (Sigma #A-1685) Wrap in foil, store at 4°C, good for years.
  • Note
   Tribromoethanol is extremely light sensitive. Its degradative products, HBr and dibromylacetaldehyde, are both very toxic.

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
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References

Relevant papers and books

If this protocol has papers or books associated with it, list those references here. Below is an example for formatting purposes. See the OpenWetWare:Biblio page for more information.

  1. Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. DOI:10.1073/pnas.78.11.6840 | PubMed ID:6947258 | HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. DOI:10.1016/s0022-2836(61)80072-7 | PubMed ID:13718526 | HubMed [Jacob-JMB-1961]
  3. ISBN:0879697164 [Ptashne-Genetic-Switch]

All Medline abstracts: PubMed | HubMed

Contact

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