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The mouse and rhinoceros fecal samples mentioned in the kit insert were collected and processed in collaboration with a zoo.
Please find below a suggested fecal extraction protocol:
1. Label 16x125mm polypropylene tubes with extraction #’s.
2. Measure 0.5g (+/- 0.05) of mixed, defrosted fecal sample into the corresponding tube, being careful to remove any litter or debris.
3. Record exact weight on fecal extraction sheet, and record any comments (unusually wet or dry, hairball).
4. Fill a labeled 12x75mm polypropylene tube with sample and freeze for future use, if required. Sample labels should include animal ID (name), sample # and date.
5. Add 5 ml of 80%EtOH in dH20 and cap tubes quickly.
6. Vortex all tubes well, making sure samples are broken up.
7. Place racked tubes in baggies in a horizontal position on a rotator and mix well for 14-18 hours (overnight).
8. Centrifuge tubes for 15 minutes at 1500 rpm.
9. Pipette 1 mL assay buffer into labeled 12x75mm polypropylene test tubes, add 1 mL, of supernatant to each corresponding tube, cap tightly and store frozen. Dilution tube labels should include animal ID (name), sample #, date and 1:10.
10. Fecals and dilutions are stored in gridded boxes. Box labels should include species, animal ID (name or #) and sex (0.1 or1.0), fecals or 1:10’s, zoo, start and finish sample # and dates.
This represents a 10-fold dilution, 5-fold in EtOH, since the original volume is 5 mL, and a further 2-fold into buffer, so that the sample will freeze.