Exercise 13.7 - Isolation of Fruiting Bodies without slugs
- 24 suspension cultures of D. discoideum
- Clinical centrifuge and tubes
- Black Millipore filters (AABP47SO)
- Cellulose pads for Millipore filters
- LPS buffer
- Cultures grown for 24-30 hours (agar or suspension) are
washed free of bacteria by several centrifugations in fresh, cold
water (2 minutes @ 300 Xg each).
- Apply the cells at a density of 6 X 10
cells per cm
to a water washed black Millipore filter supported by two cellulose
absorbent pads previously saturated with LPS Buffer.
- Allow the cells to develop in the dark at 22° C in a
humidity saturated chamber.
- Periodically withdraw a sample of the amoeba and sketch the
development of the aggregates. Note in particular the induction
of synchronous fruiting without the formation of migrating slugs.
to Table of Contents
Cell Biology Laboratory Manual
Dr. William H. Heidcamp, Biology Department, Gustavus Adolphus College,
St. Peter, MN 56082 -- email@example.com