Difference between revisions of "Performing Drosophila Crosses"

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-Collect virgin females with appropriate marker and put into a new vial
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==Preparing the Crosses==
−
-When not collecting, keep the vials from which virgins are collected at 18 degrees celsius
+
*Collect virgin females with appropriate marker and put into a new vial.
−
-The vials from which males are collected are stored at room temperature
+
*Store the vials from which virgins are collected at 18° C when not using.
−
-Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from another strain
+
*Store vials from which males are collected at room temperature.
−
-Label the vial to indicate the cross type and generation (A1 = the first generation of the first cross)
+
*Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from a different strain.
−
-Store the crosses in an incubator
+
*Label the vial to indicate the cross type and generation (A1 = the first generation of the first cross).
−
-After 5 days, flip the 14 flies into a new vial (there should be eggs visible at the surface of the food)
+
*Store the crosses in an appropriate temperature-controlled incubator.
−
-Label the vial appropriately (A2 = the second generation of the first cross)
+
*After 5 days, flip the 14 flies into a new vial (there should be eggs visible at the surface of the food).
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-Repeat this cycle until enough progeny from the crosses have been produced or until there are no parent males left
+
*Label the new vial appropriately (A2 = the second generation of the first cross).
−
- After about 10 days, the progeny from the crosses will start eclosing  
+
*Repeat this cycle until enough progeny from the crosses have been produced.
−
- 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
+
==Collecting the Progeny==
−
-Record the number of flies of each phenotype born
+
* The progeny from the crosses will start eclosing after ~10 days.
−
- Place the males and females into separate vials
+
* Record the date that the flies begin eclosing.
−
- There should be 5 - 15 progeny flies in each vial - if necessary, further separate them according to phenotype
+
* Once there are at least 20 flies in the vial, collect the progeny and sort according to phenotype.
−
- Label the vial with: Cross type/generation, gender, # of flies, phenotype, date born, and date added to vial
+
*Record the number of flies of each phenotype born.
−
- Keep the progeny vials in an incubator
+
* Place the males and females into separate vials.
−
- 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)
+
* There should be 5 - 15 progeny flies in each vial.
−
-After six days, toss the vial
+
**If necessary, separate/combine phenotypes (but not gender) to have an appropriate number of flies in the vial.
−
- Flip all progeny vials twice a week
+
* Label the vial with:  
 +
#Cross type/generation
 +
#gender
 +
# number of flies
 +
#phenotype
 +
#date born
 +
#date added to vial.
 +
* Keep the progeny vials in an appropriate 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).
 +
**Record the most likely date of birth before each sorting.
 +
*After six days, toss the vial.
 +
 
 +
==Recording Deaths of the Progeny==
 +
* Flip all progeny vials into fresh vials twice a week (every 3-4 days).
 +
*Record information on any dead flies found while flipping:
 +
#Cross type
 +
#Gender
 +
#Phenotype
 +
#Date born
 +
#Date of death

Latest revision as of 17:35, 20 June 2013

Preparing the Crosses

  • 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 appropriate temperature-controlled 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 new vial appropriately (A2 = the second generation of the first cross).
  • Repeat this cycle until enough progeny from the crosses have been produced.

Collecting the Progeny

  • The progeny from the crosses 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 (but not gender) 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 appropriate 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).
    • Record the most likely date of birth before each sorting.
  • After six days, toss the vial.

Recording Deaths of the Progeny

  • Flip all progeny vials into fresh vials twice a week (every 3-4 days).
  • Record information on any dead flies found while flipping:
  1. Cross type
  2. Gender
  3. Phenotype
  4. Date born
  5. Date of death
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