1 Fly Project Begins Objectives: Organize groups - 4 members maximum Start notebooks - Describe traits and strains, etc. Understand traits and obtain needed flies - Purebreeding Learn about handling flies - Feeding, Anesthetizing Sexing, Virgin collection Set up stock of each strain For September 18, obtain 10 virgin females of each strain (wild type, w sn m e).

2 Inheritance in Drosophila melanogaster Project Investigate genes responsible for four traits: white vs. red eyes; miniature vs. normal wings; ebony vs. tan body color, singed vs. normal bristles Determine: 1) Number of genes involved 2) If each gene is X-linked or autosomal 3) If the mutant or wild-type allele for each is dominant 4) If genes are linked or assorting independently 5) Map distances between all linked genes

3 Wild-type Drosophila melanogaster Red eyes Long wings ‘Gray/ tan’ body Smooth bristles

4 Traits Studied Miniature wings ( m ) Forked bristles ( f ) Singed bristles (sn) + sn

5 Traits Studied Scarlet eyes (st) White eyes (w) wild type (+) Brown eyes (bw)

6 Traits Studied Wild type (+) Yellow bodies (y) Ebony bodies (e)

7 Crosses to be Performed Reciprocal crosses: strain A x strain B ; strain A x strain B F1 x F1 crosses ( F1 female x F1 male ): If all X-linked or independently assorting Testcrosses ( F1 female x strain A or strain B male ): Mapping F1x pure-breeding recessive

8 Finding Information about Traits

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10 Finding Information about Traits

11 Finding Information about Traits

12 Finding Information about Traits

13 Finding Information about Traits

14 Finding Information about Traits

15 Reasons for Studying D. melanogaster Small size Short generation time (10 days - two weeks ) Easy maintenance Number of well characterized traits 13,601 protein-encoding genes identified comprehensive genetic map genome sequenced many genes comparable to humans

16 Viewing and Anesthetizing Flies Binocular dissecting microscopes ocular focus, zoom Carbon dioxide anesthetizing needles anesthetizing platforms Turn off CO 2 before leaving lab MITES!!! Wipe down everything with ethanol (70%) even when not working with flies

17 Fly Care and Maintenance Feeding - Instant Drosophila Medium Sterile vials and plugs Transfer flies to new vials with new food Stocks - Every two weeks Crosses - Every 3-4 days, until larvae in first vials Incubator 24 O C (Room temperature for now)

18 Fly Management and Supply Cleaning Morgue - unwanted adults - Freezer - unwanted cultures with potential larvae - Vial and Plug Washing - thawed vials rinsed with warm water, medium dissolved, brushed from vial, plugs rinsed with warm water until clean,

19 Sexing Flies Male Female

20 Sexing Flies Sex combs are only seen in males

21 Sexing Flies Genital arch in males

22 Sexing Flies Male Female

23 Life Cycle Pupae Eggs Larvae

24 Life Cycle

25 Virgin Collection Adults emerge from pupal cases (eclose) Males not ready to mate for 8-10 hr Females ready immediately Clear Vials before 9 AM Return within 8 hours Separate males and females into separate vials

26 Lab Supplies and Access Main Lab Tour of Fly Room Key Pad

27 Before Leaving Lab Tuesday Everyone in group must know how to: feed, transfer, anesthetize flies distinguish mutant traits from wild type collect virgins Plan - who will find information about each trait; times when each member will be in lab to clear, etc. By September 18 - Make sure you have 10 virgin females from each strain and 10 males from each strain for reciprocal crosses, which will be set up in lab.