TROPLIST: method of sacrificing xenopus for RNA or protein extraction

[Constance, Cara M.]

Dear Troplisters,
 
I am writing a protocol for approval by my institution's IACUC, and propose to extract RNA and protein from adult Xenopus tissues.  There is some contention about the most humane way to sacrifice the animals.  In the past, I have decapitated and pithed the animals before collecting the tissues that I needed.  I use this method rather than MS222 overdose to sac the animals since I want to collect the tissues immediately after the animal has expired in order to avoid degradation of the RNA or protein in the tissues.  However, I thought I would ask to see what method others use for sacrificing adult animals for tissue collection that will be subsequently used for either RNA or protein extraction.  I would appreciate knowing in particular what methods your institution's IACUCs have approved for this purpose, whether you anesthetize the animal first if using decapitation, and whether your method is considered Category C (procedures that involve no more than momentary or slight pain or distress) or Category D (procedures that may cause more than momentary or slight pain or distress for which appropriate anlgesia, anesthesia, or frequent tranquilization is provided) or Category E (Procedures that may cause pain or distress which are not relieved by anlgesia, anesthesia, or tranquilization.)  Thank you for your help!
 
Cara
 
_________________________________________________________________________
Cara M. Constance, Ph.D.
Assistant Professor
213 Colton Hall
Department of Biology
Hiram College
P.O. Box 67
11700 Hayden St.
Hiram, OH  44234
Ph: 330-569-5263
Fax: 330-569-5448
e-mail: constancecm@hiram.edu 
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