• Prepare a new culture vial with media.  Be sure the vial is kept plugged except when you are adding / removing flies.

• Select 4 - 6 mature female pupae from a wild (or mutant) subculture that has entered the hatching phase.

• Also select 4 - 6 mature male pupae (or lightly anesthetized adult males) from a mutant (or wild type) subculture.  Be sure you select males from a mutant subculture if the female pupae are wild type.  Or, if the female pupae are of the mutant type, select wild type males.

• Place the female pupae and the male pupae (or anesthetized male adults) in the newly prepared vial.  Be sure the pupae / adults are placed on the side of the vial in a dry spot.  Plug the vial.  The vial must remain on its side until the pupae hatch (1 - 3 days).

• Prepare a label for the vial and record the appropriate information for this new parental cross in your lab notebook.

• Place the label on the vial and place the vial on its side in a tray in the incubator.

• Be sure to stand the vial upright after 4 - 5 days.

• Check the vial every few days for signs of offspring larvae and, later, pupae.

• Remove the parents when the offspring pupae begin to appear.  If the parents are in good shape, they can be placed in a new vial to continue producing F₁ offspring.

• When the offspring pupae begin to hatch, sex and phenotype all adult F₁ offspring emerging during the first ten days.  Record the data in your laboratory notebook.

• Some of the F₁ offspring emerging during the first ten days of the hatch should also be used to make F₁ X F₁ crosses.
   
   

CONTACT PERSON:  Robert Weibust - weibust@mnstate.edu

LAST UPDATE:  4/25/04

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