Performing Drosophila Crosses: Difference between revisions

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-Collect virgin females with appropriate marker and put into a new vial

==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).

-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

@@ Line 1: / Line 1: @@
--Collect virgin females with appropriate marker and put into a new vial
+==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).
--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