Mouse Heart Tissue Harvest for DNA Microarrays: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
(New page: ==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. Fo...)
 
 
(8 intermediate revisions by 2 users not shown)
Line 1: Line 1:
==Overview==
==Overview==


Replace this sentence with a brief description of the protocol and its goal.
List of reagents is not yet complete


==Materials==
==Procedure==
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.
*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.


*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==
'''2.5% Avertin''' (Chem Name = Tribromoethanol, Fluka #90710)
1. Anesthesize mouse with 0.02 ml 2.5% Avertin /gram body weight.
    *Dilute 100% Avertin in warm PBS (55°C or higher)
2. Make incisions through the skin and the peritoneum and cut out approximately 100-200 mg of the tissues to be analyzed.
    *Filter sterilize, wrap in foil, store frozen aliqouts at -20°C
3. Immediately rinse the tissue in 1x PBS, blot dry, and weigh.
    *Aliqouts good for ~3 weeks at 4°C
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)
'''100% Avertin'''
     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
     *Dissolve Avertin in tert-amyl alcohol (Sigma #A-1685)  
    *Wrap in foil, store at 4°C, good for years.


100% Avertin
'''*Note:''' Tribromoethanol is extremely light sensitive. Its degradative products, HBr and dibromylacetaldehyde, are both very toxic.
    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==
==Notes==
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
#List troubleshooting tips here. 
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
#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.
No further notes are available at this time.


==References==
==References==
'''Relevant papers and books'''
'''Relevant papers and books'''
<!-- If this protocol has papers or books associated with it, list those references here.  See the [[OpenWetWare:Biblio]] page for more information. -->
 
<biblio>
No further references are available at this time
#Goldbeter-PNAS-1981 pmid=6947258
#Jacob-JMB-1961 pmid=13718526
#Ptashne-Genetic-Switch isbn=0879697164
</biblio>


==Contact==
==Contact==

Latest revision as of 14:08, 24 June 2009

Overview

List of reagents is 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

No further notes are available at this time.

References

Relevant papers and books

No further references are available at this time

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=Mouse_Heart_Tissue_Harvest_for_DNA_Microarrays&oldid=318677"