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Examining the RNAi Mechanism Kit (RNAi and Amplification; with Perishables)

SKU Code: 211392P

RNA interference (RNAi) is a technique that allows you to silence the expression of a chosen gene by specifically degrading the gene’s mRNA. The Examining the RNAi Mechanism kit allows students to use this Nobel Prize-winning technique to silence the dpy-13 gene in the non-parasitic roundworm, C. elegans. They observe wild-type worms eat a lab strain of E. coli that expresses double-stranded RNA (dsRNA) corresponding to the targeted gene, DumPY 13 (dpy-13). The ingestion of the dsRNA from the bacteria initiates the destruction of mRNA expressed from the targeted dpy-13 gene. Students examine the short, fat phenotype induced by silencing the dpy-13 gene, and then isolate DNA from the RNAi-induced dpy-13 worms, wild-type worms, and from dpy-13 worms with a deletion in the dpy-13 gene. They then amplify the region of the dpy-13 gene containing the mutation to determine if the corresponding region in the wild-type worms and the RNAi-induced dpy-13 worms has the same mutation. The deletion mutation creates a size difference in the amplified DNA fragment that can be easily observed by gel electrophoresis. In accompanying bioinformatics exercises, students explore the function of the protein encoded by the silenced gene and the relatedness of the human and worm genes. Kit includes: 0.036 g Isopropyl B-D-Thio-Galactopyranoside, 2 tubes cholesterol (5 mg/mL, 300 µL), 4 mL ampicillin, proteinase K (20 mg/mL, 25 µL), DNA marker (pBR322/BstN1), primer/loading dye mix, 4 plates dpy-13 C. elegans, 4 plates wild-type (N2) C. elegans, E. coli slants (1 OP50 and 1 dpy-13 RNAi feeding strain), 25 PCR beads (in 0.2-mL tubes), lysis buffer (0.5-mL tube), 1 mL sterile distilled water, 6 sterile dropping pipets, 2 melt n’ pour NGM lite media bottles (100 mL and 300 mL), 40 petri dishes (60 mm), 3 disposable inoculating loops, 3 sterile culture tubes, 3 LB broth (3-mL vials), 5 mL mineral oil, 8 glass tubes of sterile toothpicks, and instructor’s manual with reproducible student guide.
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Manufacturer: Carolina
Department: Biotechnology
Product Line: STOCK
Weight: Width: 4.6 LBS