2.2.5. Mesh bags
Mesh bags are sturdy plastic bags with a small pore fine mesh inside. The sample is placed on one side of the mesh, ground from the outside and the homogenate is collected from the other side of the mesh, filtering out large particles.
- Pros: Medium-large volume; easy; cheap; no cross-contamination.
- Cons: Lack of
uniformity; split bags.
- Place up to 30 frozen bees in a disposable mesh-bag (e.g., www.bioreba.com; #430100).
- Add 500 μl buffer per bee.
- Flash-freeze the entire bag in liquid nitrogen.
- Remove from liquid nitrogen.
- Wait 30 seconds.
- Pulverize contents by grinding the bag with a large pestle for 2 minutes on a hard surface, taking care not to damage the bag.
- Massage the bag until completely thawed.
- Remove one (or more) 100 μl aliquots of homogenate.
- As an alternate method, described in section ## samples can be crushed in disposable mesh bags using a heavy rolling pin.