BISC219/F11:RNA interference: Difference between revisions
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[[BISC219/F11: RNAi Lab 9 | Lab 9: Induction of bacteria for RNAi]]<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 10 | Lab 10: Scoring your worms ]]<br> | ||
[[BISC219/F11: RNAi Lab 11 | Lab 11: ]]<br><br> | [[BISC219/F11: RNAi Lab 11 | Lab 11: Individual Writing Conferences]]<br><br> | ||
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| | |Sign up for a time to meet and discuss your final paper draft with your instructor - we will not meet in lab this week. | ||
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Latest revision as of 09:02, 8 November 2011
These labs were developed with the help of the Silencing Genomes project of the Dolan DNA Learning Center.
RNAi General Information
Media Recipes
Lab 5: Picking your gene to RNAi
Lab 6: Cloning your gene of interest
Lab 7: Picking your transformant
Lab 8: Plasmid purification and transformation
Lab 9: Induction of bacteria for RNAi
Lab 10: Scoring your worms
Lab 11: Individual Writing Conferences
Schedule of Experiments
Lab # | Dates | Activity | Outside lab time |
---|---|---|---|
5 | 9/29 - 10/5 | Pick a gene of interest Set up a PCR reaction to clone the gene |
Examine the results of the agarose gel electrophoresis |
6 | 10/12 - 10/18 | Restriction enzyme digest of PCR product Cleanup and ligation into the pPD129.36 vector |
The day after lab: Check control and transformation plates for growth - save your transformation plate |
7 | 10/19 - 10/25 | Colony PCR to check for transformation | The night before next lab: Set up an overnight culture of a single colony from your transformation |
8 | 11/2 - 11/8 | Plasmid isolation from the BL21 cells Quantification of DNA |
The day after lab: Check control and transformation plates for growth - save your transformation plate |
9 | 11/9 - 11/15 | Induction of the bacteria to produce RNA Seed plates and dry for bacterial feeding RNAi |
4 days later: Pick 2 L4 hermaphrodite worms of N2 and rrf-3 genotype to 2 RNAi plates for each genotype and make 1 control "mock" plate for each genotype as well (6 plates total) |
10 | 11/16 - 11/22 | Examine the phenotypes of the fed worms - compare to control N2 worms and worms containing a mutation in the gene you are examining Collection of treated worms and RNA purification |
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11 | 11/28 - 12/2 | Sign up for a time to meet and discuss your final paper draft with your instructor - we will not meet in lab this week. |