BISC219/F12:RNA interference

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These labs were modified from the work of Janet Andersen, Alexander Krichevsky, Joerg R. Leheste, and Daniel J. Moloney at the Dept. of Biochemistry and Cellular BIology at Stony Brook University with the Silencing Genomes project of the Dolan DNA Learning Center in Cold Spring Harbour, New York. providing some of the worm strains.

Background Information on Project 3: Investigating Gene Function & Regulation Using RNAi
Media Recipes
Lab 7: Identifying a bacterial colony containing our plasmid of interest
Lab 8: Creating the feeding strain of bacteria for RNAi
Lab 9: Induction of feeding strain to produce dsRNA and feeding worms
Lab 10: Phenotypic analysis of treated vs untreated worms
Lab 11: Data Analysis Workshop
Lab 12: Optional Conferences

Schedule of Experiments

Lab # Dates Activity Outside lab time
7 10/23 - 10/29 Picking a colony of bacteria containing the RNAi plasmid

PCR to confirm presence of gene of interest

The night before next lab: set up an overnight culture of your colony of interest
8 10/30 - 11/5 Miniprep of plasmid and transformation into feeding bacteria strain The day after lab: check your transformed bacteria for colony growth

The night before next lab: set up an overnight culture of your colony of interest

9 11/7 - 11/13 Induction of dsRNA production, seeding of RNAi plates and addition of worms
10 11/14 - 11/20 Chemotaxis Assay
11 11/26 - 11/30 Writing Workshop
12 12/3 - 12/7 Writing Conferences