BISC 219/F10: Lab 2 Mutant Hunt: Difference between revisions
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== Mutant Hunting == | == Mutant Hunting == | ||
'''To Do Today (per group)'''<br> | '''To Do Today (per group)'''<br> | ||
#Choose one of the six plates at the front of the room <br> | #Choose one of the six plates of "mutagenized" worms at the front of the room <br> | ||
#Scan | #Scan the “mutagenized” worms on this plate for visible dumpy mutants | ||
#Transfer 1 putative mutant hermaphrodite to each of three new plates. Take care not to transfer any other animals | #Transfer 1 putative dpy mutant hermaphrodite to each of three new plates. Take care not to transfer any other animals or eggs other than your putative mutant.<br> | ||
#Label these identical plates with the hermaphrodite mutant's strain information and your group's information. Use your <font color= blue><b>BLUE </b></font color> Sharpie for labeling | #Label these 3 identical plates with the hermaphrodite mutant's strain information and your group's information (team color, initials, lab day, date). Use your <font color= blue><b>BLUE </b></font color> Sharpie for labeling these linkage analysis plates. | ||
#Put an elastic around the three | #Put an elastic around the three linkage determination plates. Place them in your group's plastic box (make sure your box is labeled with your names and lab section on a piece of your team color tape). | ||
#Place your box on the shelf in the incubator for your lab section.<br> | #Place your box on the shelf in the incubator for your lab section.<br> | ||
#Incubate your worms at 23°C for 3 days.<br> | #Incubate your worms at 23°C for 3 days.<br> | ||
<br> | <br> | ||
'''3 Days After Lab'''<br> | '''3 Days After Lab'''<br> | ||
#Examine your three plates containing your dpy worms from the mutant hunt. Are all the worms on the plates the dpy phenotype? If '''YES''', then start a cross between 4 dpy L4 hermaphrodites (still have the clearing about 1/2 way down the body of the worm) and 3-4 wild type male worms. | #Examine your three plates containing your dpy worms from the mutant hunt and their F1 progeny. Are all the worms on the plates the dpy phenotype? If '''YES''', then start a cross between 4 of these F1 dpy L4 hermaphrodites (should still have the vulval clearing about 1/2 way down the body of the worm) and 3-4 wild type male worms. You will have to search to find the N2 male worms among the hermaphrodites in the N2 plates provided on the bench on the right side near the sink. Note that if you don't have all dumpy mutants in the F1 generation of your cross, you should not use that plate to select worms for the next cross. If none of your 3 plates contains all dumpy mutants email your instructor ASAP!<br> | ||
#Set up two identical mating plates | # If you have an appropriate plate for continuing the next cross: Set up two identical mating plates with 3-4 L4 hermaphrodites and 3-4 wild type males.<br> | ||
#Label these identical plates with the hermaphrodite mutant's strain information and your group's information. Use your <font color= blue><b>BLUE </b></font color> Sharpie for labeling mapping | #Label these identical plates with the hermaphrodite mutant's strain information and your group's information including the date. Use your <font color= blue><b>BLUE </b></font color> Sharpie for labeling these linkage analysis/mapping plates. <br> | ||
#Incubate your worms at 23°C until next lab period. | #Incubate your worms at 23°C until next lab period. | ||
<br><br> | <br><br> | ||
Revision as of 07:35, 2 September 2010
Mutant Hunting
To Do Today (per group)
- Choose one of the six plates of "mutagenized" worms at the front of the room
- Scan the “mutagenized” worms on this plate for visible dumpy mutants
- Transfer 1 putative dpy mutant hermaphrodite to each of three new plates. Take care not to transfer any other animals or eggs other than your putative mutant.
- Label these 3 identical plates with the hermaphrodite mutant's strain information and your group's information (team color, initials, lab day, date). Use your BLUE Sharpie for labeling these linkage analysis plates.
- Put an elastic around the three linkage determination plates. Place them in your group's plastic box (make sure your box is labeled with your names and lab section on a piece of your team color tape).
- Place your box on the shelf in the incubator for your lab section.
- Incubate your worms at 23°C for 3 days.
3 Days After Lab
- Examine your three plates containing your dpy worms from the mutant hunt and their F1 progeny. Are all the worms on the plates the dpy phenotype? If YES, then start a cross between 4 of these F1 dpy L4 hermaphrodites (should still have the vulval clearing about 1/2 way down the body of the worm) and 3-4 wild type male worms. You will have to search to find the N2 male worms among the hermaphrodites in the N2 plates provided on the bench on the right side near the sink. Note that if you don't have all dumpy mutants in the F1 generation of your cross, you should not use that plate to select worms for the next cross. If none of your 3 plates contains all dumpy mutants email your instructor ASAP!
- If you have an appropriate plate for continuing the next cross: Set up two identical mating plates with 3-4 L4 hermaphrodites and 3-4 wild type males.
- Label these identical plates with the hermaphrodite mutant's strain information and your group's information including the date. Use your BLUE Sharpie for labeling these linkage analysis/mapping plates.
- Incubate your worms at 23°C until next lab period.
