BISC 219/2009:RNA interference: Difference between revisions
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{{Template:BISC 219/2009}} | {{Template:BISC 219/2009}} | ||
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These labs were developed with the help of the '''Silencing Genomes''' project of the Dolan DNA Learning Center.<br><br> | |||
[[BISC 219: Mod 2 RNAi General Information| RNAi General Information]] <br> | [[BISC 219: Mod 2 RNAi General Information| RNAi General Information]] <br> | ||
[[BISC 219: Mod 2 Media Recipes | Media Recipes]]<br> | [[BISC 219: Mod 2 Media Recipes | Media Recipes]]<br> | ||
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[[BISC 219: Mod 2 Lab 8 | Lab 8: PCR Reaction Cleanup and Sequencing]]<br><br> | [[BISC 219: Mod 2 Lab 8 | Lab 8: PCR Reaction Cleanup and Sequencing]]<br><br> | ||
=='''Schedule of Experiments'''== | =='''Schedule of Experiments'''== |
Revision as of 17:02, 20 September 2009
These labs were developed with the help of the Silencing Genomes project of the Dolan DNA Learning Center.
RNAi General Information
Media Recipes
Lab 4: Picking your gene to RNAi
Lab 5: Plasmid DNA isolation and transformation
Lab 6: Induction of RNAi plasmid and C. elegans feeding
Lab 7: Single Worm PCR and Agarose Gel Electrophoresis
Lab 8: PCR Reaction Cleanup and Sequencing
Schedule of Experiments
Lab # | Dates | Activity | Outside lab time |
---|---|---|---|
4 | 9/29 - 10/5 | Pick a single bacterial colony containing a plasmid for RNAi | The night before next lab: Set up an overnight culture of your colony |
5 | 10/6 - 10/14 | Plasmid DNA isolation Transformation of the isolated plasmid into the HT115(DE) feeding strain |
The next day: Check control and transformation plates for growth - save your transformation plate |
6 | 10/19 - 10/23 | Induction of the bacteria to produce RNA Seed plates and dry for bacterial feeding RNAi |
3 days later: Pick 3 gravid adult worms of N2, rrf-3 and rde-1 genotype to 2 RNAi plates for each genotype (6 plates total) |
7 | 10/26 - 10/30 | Examine the phenotypes of the fed worms - compare to control N2 worms and worms containing a mutation in the gene you are examining Single worm PCR of N2 and RNAi treated worms |
|
8 | 11/4 - 11/10 | Clean up PCR reactions with Qiagen Min-Elute kit Run sequencing reaction |
Analyze your sequencing data |