Examining the RNAi Mechanism Kit (RNAi, Amplification, and Electrophoresis Kit with CarolinaBLU®; with voucher)

Product Code: 211393

Description

Advanced– For experienced high school and college classes; requires some basic 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.

Kit includes a voucher to request perishable materials later at your convenience. Contact us or return the voucher to request delivery of perishables. Kit also includes FREE 1-year access to digital resources that support 3-dimensional instruction for NGSS.

 

Includes

  • 1 Ampicillin, 10 mg/mL, 4 mL (voucher item)
  • 2 Cholesterol, 5 mg/mL, 0.3 mL (voucher item)
  • 1 IPTG, 0.036 g (voucher item)
  • 1 Proteinase K, 20 mg/mL, 25 μL (voucher item)
  • 1 DNA Marker pBR322/BstN1, 200 μL (voucher item)
  • 4 dpy-13 (e458) Worms on NGM-lite Plates (voucher item)
  • 1 dpy-13 Primer/Loading Dye Mix, 700 μL (voucher item)
  • 1 E. coli RNA Feeding Strain (dpy-13) LB Amp/Tet Slant (voucher item)
  • 1 E. coli OP50 on LB Slant (voucher item)
  • 4 Wild-type Worms on NGM-lite Plates (voucher 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