Difference between revisions of "Performing Drosophila Crosses"
From Bridges Lab Protocols
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| − | + | *Collect virgin females with appropriate marker and put into a new vial. | |
| − | + | *Store the vials from which virgins are collected at 18° C when not using. | |
| − | + | *Store vials from which males are collected at room temperature. | |
| − | + | *Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from a different strain. | |
| − | + | *Label the vial to indicate the cross type and generation (A1 = the first generation of the first cross). | |
| − | + | *Store the crosses in an incubator. | |
| − | + | *After 5 days, flip the 14 flies into a new vial (there should be eggs visible at the surface of the food). | |
| − | + | *Label the vial appropriately (A2 = the second generation of the first cross). | |
| − | + | *Repeat this cycle until enough progeny from the crosses have been produced or until there are no parent males left. | |
| − | + | * The progeny will start eclosing after ~10 days. | |
| − | + | * Record the date that the flies begin eclosing. | |
| − | + | * Once there are at least 20 flies in the vial, collect the progeny and sort according to phenotype. | |
| − | + | *Record the number of flies of each phenotype born. | |
| − | + | * Place the males and females into separate vials. | |
| − | + | * There should be 5 - 15 progeny flies in each vial. | |
| − | + | **If necessary, separate/combine phenotypes to have an appropriate number of flies in the vial. | |
| − | + | * Label the vial with: | |
| − | + | #Cross type/generation | |
| − | + | #gender | |
| − | + | # number of flies | |
| + | #phenotype | ||
| + | #date born | ||
| + | #date added to vial. | ||
| + | * Keep the progeny vials in an incubator. | ||
| + | * Repeat this process during the six days following the date of first eclosure | ||
| + | **Wait to sort until there are at least 20 flies in the vial (usually 2 to 3 times per vial) | ||
| + | *After six days, toss the vial | ||
| + | * Flip all progeny vials twice a week | ||
Revision as of 18:00, 19 June 2013
- Collect virgin females with appropriate marker and put into a new vial.
- Store the vials from which virgins are collected at 18° C when not using.
- Store vials from which males are collected at room temperature.
- Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from a different strain.
- Label the vial to indicate the cross type and generation (A1 = the first generation of the first cross).
- Store the crosses in an incubator.
- After 5 days, flip the 14 flies into a new vial (there should be eggs visible at the surface of the food).
- Label the vial appropriately (A2 = the second generation of the first cross).
- Repeat this cycle until enough progeny from the crosses have been produced or until there are no parent males left.
- The progeny will start eclosing after ~10 days.
- Record the date that the flies begin eclosing.
- Once there are at least 20 flies in the vial, collect the progeny and sort according to phenotype.
- Record the number of flies of each phenotype born.
- Place the males and females into separate vials.
- There should be 5 - 15 progeny flies in each vial.
- If necessary, separate/combine phenotypes to have an appropriate number of flies in the vial.
- Label the vial with:
- Cross type/generation
- gender
- number of flies
- phenotype
- date born
- date added to vial.
- Keep the progeny vials in an incubator.
- Repeat this process during the six days following the date of first eclosure
- Wait to sort until there are at least 20 flies in the vial (usually 2 to 3 times per vial)
- After six days, toss the vial
- Flip all progeny vials twice a week
