Bridges Lab Protocols - User contributions [en]

Performing Drosophila Crosses

2013-06-20T17:35:16Z

IsabelleHatfield:

==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:
#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

Performing Drosophila Crosses

2013-06-19T18:00:26Z

IsabelleHatfield:

*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

Performing Drosophila Crosses

2013-06-19T17:39:20Z

IsabelleHatfield: 1st draft

-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
-The vials from which males are collected are stored at room temperature
-Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from another 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
- After about 10 days, the progeny from the crosses will start 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
-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, further separate them according to phenotype
- Label the vial with: Cross type/generation, gender, # of flies, phenotype, date born, and date added to vial
- Keep the progeny vials in an incubator
- 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)
-After six days, toss the vial
- Flip all progeny vials twice a week