BISC219/F11: RNAi Lab 10: Difference between revisions
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'''B.''' Plate wild type ''C. elegans'' worms (N2 and ''rrf-3'' strains) on feeder plates made as described (containing bacteria expressing dsRNA of our gene of interest). <BR> | '''B.''' Plate wild type ''C. elegans'' worms (N2 and ''rrf-3'' strains) on feeder plates made as described (containing bacteria expressing dsRNA of our gene of interest). <BR> | ||
'''C.''' Observe phenotype change in progeny caused by RNAi silencing or knockdown of the gene of interest compared to control worms of same strains that were NOT fed feeder strain bacteria.<BR> | '''C.''' Observe phenotype change in progeny caused by RNAi silencing or knockdown of the gene of interest compared to control worms of same strains that were NOT fed feeder strain bacteria.<BR> | ||
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==Links to Labs& Project Info== | |||
'''Series1:'''<BR> | |||
[[BISC219/F11: Worm Info | Worm Info]] <br> | |||
[[BISC219/F11: Gene Linkage | Lab 1: Worm Boot Camp & Sex-Linked or Autosomal Start]]<BR> | |||
[[BISC219/F11: Lab 2 | Lab 2: Sex-Linked or Autosomal Finale]]<br> | |||
'''Series2:'''<BR> | |||
[[BISC219/F11: Gene Mapping Info | Background: Classical Forward Genetics and Gene Mapping]]<br> | |||
[[BISC219/F11: Lab 2 Mutant Hunt | Lab 2: Mutant Hunt]]<br> | |||
[[BISC219/F11: Lab 3 | Lab 3: Linkage Test Part 1]]<br> | |||
[[BISC219/F11: Lab 4 | Lab 4: Linkage Test Part 2, Mapping and Complementation]]<br> | |||
[[BISC219/F11: Lab 5 | Lab 5: Finish Complementation; Mapping Continued]]<br> | |||
[[BISC219/F11: Lab 6 | Lab 6: DNA sequence analysis; Mapping Continued]]<BR> | |||
[[BISC219/F11: Lab 7 | Lab 7: Complete Mapping: Score]]<br> | |||
'''Series3:'''<BR> | |||
[[BISC219/F11:RNA interference | RNA interference]]<BR> | |||
[[BISC219/F11: RNAi General Information | RNAi General Information]] <br> | |||
[[BISC219/F11: Media Recipes | Media Recipes]]<br> | |||
[[BISC219/F11: RNAi Lab 5 | Lab 5: Picking your gene to RNAi]]<br> | |||
[[BISC219/F11: RNAi Lab 6 | Lab 6: Cloning your gene of interest]]<br> | |||
[[BISC219/F11: RNAi Lab 7 | Lab 7: Picking your transformant]]<br> | |||
[[BISC219/F11: RNAi Lab 8 | Lab 8: Plasmid purification and transformation]]<br> | |||
[[BISC219/F11: RNAi Lab 9 | Lab 9: Induction of bacteria for RNAi]]<br> | |||
[[BISC219/F11: RNAi Lab 10 | Lab 10: Scoring your worms]]<br> | |||
[[BISC219/F11: RNAi Lab 11 | Lab 11: ]]<br><br> | |||
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Revision as of 10:23, 17 May 2011
Series 3 Reverse Genetics: Scoring RNAi Worms
3 days before next lab:
- Come into lab and find your stack of plates.
- On 2 of the experimental plates add 2 L4 wild type (N2) hermaphrodites
- On 2 of the experimental plates add 2 L4 rrf-3 hermaphrodites
- On 1 of the control plates add 2 L4 wild type (N2) hermaphrodites
- On 1 of the control plates add 2 L4 rrf-3 hermaphrodites
- Wrap all of your plates in an elastic and stick in your lab day box in the worm incubator set at 23°C
Phenotypic Analysis
Today in lab you will examine your RNAi worms and their progeny.
- Examine worms with a known bli-1 mutation to see what knockdown looks like.
- Compare your RNAi worms to worms that have a known bli-1 mutation - how do they compare?
- Examine your control worms to review wild type phenotype in comparison to mutants
- Compare your RNAi worms to the control worms - are they the same phenotype? Different? What do the “control” worms tell you?
- Examine your RNAi’d wild type (N2) and then your RNAi’d rrf-3 worms. Do you see any differences between the two treated strains of worms?
Be sure to record all of your results in your lab notebook.
Take pictures of your control and RNAi worms to use in the results section of your next paper.
Assignment
Remember to check the Assignment section of the wiki for instructions about the graded assignment due in the next lab and check the Weekly Calendar for other work to accomplish before the next lab.