Performing Drosophila Crosses: Difference between revisions

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-Collect virgin females with appropriate marker and put into a new vial
*Collect virgin females with appropriate marker and put into a new vial.
-When not collecting, keep the vials from which virgins are collected at 18 degrees celsius
*Store the vials from which virgins are collected at 18° C when not using.
-The vials from which males are collected are stored at room temperature
*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 another strain
*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)
*Label the vial to indicate the cross type and generation (A1 = the first generation of the first cross).
-Store the crosses in an incubator
*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)
*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)
*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
*Repeat this cycle until enough progeny from the crosses have been produced or until there are no parent males left.
- After about 10 days, the progeny from the crosses will start eclosing  
* The progeny will start eclosing after ~10 days.
- Record the date that the flies begin eclosing
* 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
* 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
*Record the number of flies of each phenotype born.
- Place the males and females into separate vials
* Place the males and females into separate vials.
- There should be 5 - 15 progeny flies in each vial - if necessary, further separate them according to phenotype
* There should be 5 - 15 progeny flies in each vial.
- Label the vial with: Cross type/generation, gender, # of flies, phenotype, date born, and date added to vial
**If necessary, separate/combine phenotypes to have an appropriate number of flies in the vial.
- Keep the progeny vials in an incubator
* Label the vial with:  
- Repeat this process during the six days following the date they first eclosed, waiting each time until there are at least 20 flies in the vial (usually 2 to 3 times per vial)
#Cross type/generation
-After six days, toss the vial
#gender
- Flip all progeny vials twice a week
# 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:
  1. Cross type/generation
  2. gender
  3. number of flies
  4. phenotype
  5. date born
  6. 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