Pancreatic RNA extraction
1. Prepare liquid nitrogen, a middle-size motor and a pestle, QIAshredder, 2-mL
tubes, and RNeasy mini kit. Add -ME to RLT buffer.
2. Excise pancreas (--control the size to be relatively small*) and immediately place
it in a motor that contains liquid nitrogen.
3. Let the tissue freeze completely and grind it into small pieces. Don’t allow it to
be thawed.
4. Pour the pieces with a small amount of liquid nitrogen directly into a 2-mL tube.
NEVER close the cap!
5. Immediately after liquid nitrogen evaporates, add 600l RLT buffer and transfer
the mix to a QIAshredder column.
6. Spin at a max speed for 2 min at room temperature.
7. Place the tube on ice until all samples are prepared up to this step.
8. For the rest of procedure, follow the manufacturer’s protocol (RNeasy kit).
* Up to about 14-16 days old, the whole pancreas may be used. For older mice, trim
the tissue.