Description
Advanced—For experienced high school and college classes; requires some technical skill.
RNA interference (RNAi) is a technique that allows you to silence expression of a chosen gene by degrading the gene’s mRNA. This kit lets students use the 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). Ingestion of dsRNA from bacteria initiates destruction of mRNA expressed from the targeted dpy-13 gene. Students examine the short, fat phenotype induced by silencing the dpy-13 gene, 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 amplify the region of the dpy-13 gene with the mutation to see if the corresponding region in the wild-type worms and RNAi-induced dpy-13 worms has the same mutation. The deletion mutation creates a size difference in the amplified DNA fragment, easily observed by gel electrophoresis. In bioinformatics exercises, they explore function of protein encoded by the silenced gene and relatedness of human and worm genes.
Product Features
- Demonstrate to your students the power of silencing a single gene
- Teach about a powerful method for determining gene function
- Explore the relationship between phenotype, genotype, and RNAi-induced phenotype
- Perform bioinformatics exercises exploring protein function and C. elegans, and human relatedness
- Comes with safe-to-use CarolinaBLU® stain
- Includes FREE 1-year access to Carolina Science Online® digital resources
Note: Kit includes the perishable materials when shipped.
Includes
- 1 Ampicillin, 10 mg/mL, 4 mL (perishable item)
- 2 Cholesterol, 5 mg/mL, 0.3 mL (perishable item)
- 1 IPTG, 0.036 g (perishable item)
- 1 Proteinase K, 20 mg/mL, 25 μL (perishable item)
- 1 DNA Marker pBR322/BstN1, 200 μL (perishable item)
- 4 dpy-13 (e458) Worms on NGM-lite Plates (perishable item)
- 1 dpy-13 Primer/Loading Dye Mix, 700 μL (perishable item)
- 1 E. coli RNA Feeding Strain (dpy-13) LB Amp/Tet Slant (perishable item)
- 1 E. coli OP50 on LB Slant (perishable item)
- 4 Wild-type Worms on NGM-lite Plates (perishable item)
- 1 1x Lysis Buffer, 0.5 mL
- 6 Sterile Bulb Pipets
- 3 Sterile Culture Tubes, 15 mL
- 3 Sterile Inoculating Loops
- 3 LB Broth, 3 mL
- 1 Mineral Oil, 5 mL
- 1 Melt ‘n Pour NGM-lite, 300 mL
- 1 Melt ‘n Pour NGM-lite, 100 mL
- 40 Sterile Petri Dishes, 60 mm
- 25 Ready-to-Go™ PCR Beads (0.2 mL tubes)
- 1 Sterile Distilled Water, 1 mL
- 8 Sterile Toothpicks
- 1 Agarose, 6 g
- 4 Gloves
- 1 CarolinaBLU&trade Gel and Buffer Stain, 7 mL
- 1 CarolinaBLU&trade Final Stain 250 mL
- 8 Staining Trays
- 1 TBE Buffer, 200 mL
- 1 Carolina Science Online® Resources (1-year subscription)
- 1 Teacher’s Manual and Reproducible Student Guide