Davebrid: wrote initial protocol based on Broughton et al

2012-11-30T14:51:50Z

wrote initial protocol based on Broughton et al

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[[ Category: Drosophila ]]
[[ Category: Genotyping ]]
[[ Category: Aging]]

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==Generating Experimental Flies==

===Setting up Tissue Specific shRNA Expression Experiment===

# Generate a large amount of the driver line, since you will need these as both controls and one for each shRNA being tested. See [[ Maintenance of Fly Stocks ]] for details.
# Order and propagate the shRNA lines.

====Parental Crosses====
# Set up parental cross of Driver/+ vs shRNA/+. If either the driver or the shRNA is homozygous then, you will not get all of the potential controls, but the experiment will still work as long of the alleles is heterozygous.
# Place 10 Females with 3-10 Male flies in a parental cross vial.
# To determine how many parental crosses are needed presume that 10 females will generate '''XXX''' flies and calculate backwards.
# After ~10 days remove parental flies to prevent mixing of parental with progenies.

====Analysis of Progeny====
# Collect eclosed progeny over a 8h period after they have started hatching.
# Separate progeny based on gender and genotype, placing 10 flies in a vial. The goal should be 150-250 flies per genotype/gender
# Transfer flies to a new vial twice weekly under anaesthesia maintaining 10 flies per vial.
# Check for deaths 5-6 times per week.

==References==

* [http://www.pnas.org/content/102/8/3105.full.pdf+html Longer lifespan, altered metabolism, and stress resistance in Drosophila from ablation of cells making insulin-like ligands]

Measuring Lifespan in Drosophila - Revision history

Davebrid: wrote initial protocol based on Broughton et al