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

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Examining the RNAi Mechanism Kit, RNAi, Amplification, and Electrophoresis Kit with CarolinaBLU. RNA interference (RNAi) is a technique that allows you to silence expression of a chosen gene by degrading the gene's mRNA. Kit lets students use the Nobel Prize-winning technique to silence the dpy-13 gene in 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 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 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 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: 0.036g Isopropyl B-D-Thio-Galactopyranoside, 2 tubes cholesterol (5mg/mL, 300µL), 4 mL ampicillin, proteinase K (20mg/mL, 25µL), 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), 1mL sterile distilled water, 6 sterile dropping pipets, 2 melt n’ pPour NGM lite media bottles (100mL and 300mL), 40 petri dishes (60mm), 3 disposable inoculating loops, 3 sterile culture tubes, 3 LB broth (3-mL vials), 5mL mineral oil, 8 glass tubes of sterile toothpicks, 8g agarose, 20x TBE (200 mL), 8 staining trays, CarolinaBlu, and instructor’s manual with reproducible student guide.

FREE 1-year access to digital resources that support NGSS instruction included.


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What’s Included:
  • 1 Ampicillin, 10 mg/mL, 4 mL  
  • 2 Cholesterol, 5 mg/mL, 0.3 mL  
  • 1 IPTG, 0.036 g  
  • 1 Proteinase K, 20 mg/mL, 25 μL  
  • 1 DNA Marker pBR322/BstN1, 200 μL  
  • dpy-13 (e458) Worms on NGM-lite Plates  
  • dpy-13 Primer/Loading Dye Mix, 700 μL  
  • E. coli RNA Feeding Strain (dpy-13) LB Amp/Tet Slant  
  • E. coli OP50 on LB Slant  
  • 4 Wild-type Worms on NGM-lite Plates  
  • 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
  • CarolinaBLU&trade Gel and Buffer Stain, 7 mL
  • CarolinaBLU&trade Final Stain 250 mL
  • 8 Staining Trays
  • 1 TBE Buffer, 200 mL
  • 1 Teacher's Manual and Reproducible Student Guide
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Needed But Not Included:
  •  Document Camera or Overhead Projector
  •  Stereomicroscope
  •  Bunsen Burner
  •  Containers with Cracked or Crushed Ice
  •  Container with Liquid Nitrogen or Dry Ice
  •  Digital or Instant Camera (optional)
  •  Ethanol, 95%
  •  Gel Casting Trays and Combs
  •  Gel Electrophoresis Chamber
  •  Masking Tape
  •  Microcentrifuge
  •  Microcentrifuge Adaptors
  •  Micropipettor and Tips, 1-20 μL and 20-200 μL
  •  Microcentrifuge Tube Rack
  •  Mineral Oil
  •  PCR Tubes, 0.2 or 0.5 μL
  •  Permanent Markers, Black and Red
  •  Power Supply
  •  Steel Scalpel, Spatula, or Forceps
  •  Thermal Cycler
  •  UV or White Light Transilluminator
  •  Water Bath, 60°C (for agarose solution)
  •  Heavy-duty Plastic Bag
  •  Household Bleach
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