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Synaptic Proteins: Protocols

Welcome to the Home of Vesicle Trafficking

A small repository of synaptic protein information

Protocols

Caution! In converting these protocols to HTML, some units have been scrambled.
Specifically, microliters (ul) often appear as milliliters (ml). Think first!


DROSOPHILA HEAD COLLECTION
(Baker Lab)

Equipment

flies frozen in 250 ml centrifuge bottles
sieves: 106, 355, 600 and 850 mm
paintbrush
collection tubes
funnel
dry ice
jacket, hat and gloves!


Preparation

1. Freeze flies in 250 ml centrifuge bottle(s) @ -80oC (30').
2. Cool sieves @ -20oC for 20' prior to use.


Procedure

1. Arrange sieves as follows (top to bottom):
parts retained
850 mm bodies
600 small bodies + some heads
355 heads
106 fly dust, legs

2. Shake flies with moderate strength 4-6X for 3" each time.

3. Pour flies onto top sieve.

4. Brush gently so as not to jam flies into mesh openings. Females tend to be a little larger than males; you may want to pass the 600mm material back over the 850 mm sieve 2- 4X.

5. Collect heads into tared tubes. Keep frozen until used. Obtain a weight (?).
acc 1/90


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