Mouse Heart Tissue Harvest for DNA Microarrays

Overview

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Materials

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• 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

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

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References

Relevant papers and books

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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 |
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 |
3. ISBN:0879697164

Contact

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