Wednesday, June 25, 2014

Setting up for a hemiclone assay

 Several of the assays we are conducting in the lab this summer are examining the genetic basis of female mating behaviours. There are many ways to do these assays, but (in my opinion) one of the best ways to measure ofstanding genetic is using hemiclonal analysis. Both Mireille and Arnold (above) have been learning the ropes of this technique, and this week set up the first (of many) assays.

The first step involves mating clone males (which carry a randomly-sampled haploid genome, and set of translocated autosomal chromosomes) with many wild-type virgin female collected from one of our outbred populations. These mated flies are placed into half-pint collectors outfiltted with 35mm (diameter) petrie dishes containing a grape-juice/agar media for no more than 18h.

During that time, eggs are laid on the surface (see above).
As fruit fly development can be strongly influenced by variation in larval density, it is essential that our assay vials are standardized. This is done by counting exact numbers of eggs (under the microscope, using a paintbrush)...
...which are then gently cut from the surface of the juice "cookie"....
...and transferred to vials containing ~10ml of our lab's standard banana Drosophila media!
Due to the nature of the cross between the clone males and the wild-type females, there is a 50% mortality (due to chromosomal imbalances). Thus, for every 100 eggs we transfer into the vial, only 50 will hatch into larvae. To further establish standard developmental conditions in the vials, we add an additional 50 eggs from one of our other populations that carry a recessive-brown-eyed marker, yielding a total of ~100 larvae per vial (which mimics the typical developmental conditions experienced in the our lab populations).

Now that our vials are set up, we'll incubate them, and then collect hemiclonal (red-eyed) females as they eclose starting 9d later. Check back for updates!

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