Performing Drosophila Crosses - Revision history

IsabelleHatfield at 17:35, 20 June 2013

2013-06-20T17:35:16Z

← Older revision		Revision as of 17:35, 20 June 2013
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			==Preparing the Crosses==
	*Collect virgin females with appropriate marker and put into a new vial.		*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 the vials from which virgins are collected at 18° C when not using.
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	*Once 10 virgin females from a strain have been collected, add 4 males with the appropriate marker from a different 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 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).		*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 new 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.
	* The progeny will start eclosing after ~10 days.
			==Collecting the Progeny==
			* The progeny from the crosses 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.
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	* 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.		* 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.		**If necessary, separate/combine phenotypes (but not gender) to have an appropriate number of flies in the vial.
	* Label the vial with:		* Label the vial with:
	#Cross type/generation		#Cross type/generation
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	#date born		#date born
	#date added to vial.		#date added to vial.
	* Keep the progeny vials in an incubator.		* Keep the progeny vials in an appropriate incubator.
	* Repeat this process during the six days following the date of first eclosure		* 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)		**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		**Record the most likely date of birth before each sorting.
	* Flip all progeny vials twice a week		*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

IsabelleHatfield at 18:00, 19 June 2013

2013-06-19T18:00:26Z

← Older revision		Revision as of 18:00, 19 June 2013
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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

IsabelleHatfield: 1st draft

2013-06-19T17:39:20Z

1st draft

New page

-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