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Students use RNAi 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. Comes with perishable materials.
RNA interference (RNAi) is a technique that allows you to silence the expression of a chosen gene by specifically degrading the gene‚Äôs 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‚Äô 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‚Äôs manual with reproducible student guide.
|Ampicillin, 10 mg/mL, 4 mL||1||Included|
|Cholesterol, 5 mg/mL, 0.3 mL||2||Included|
|IPTG, 0.036 g||1||Included|
|Proteinase K, 20 mg/mL, 25 μL||1||Included|
|DNA Marker pBR322/BstN1, 200 μL||1||Included|
|dpy-13 (e458) Worms on NGM-lite Plates||4||Included|
|dpy-13 Primer/Loading Dye Mix, 700 μL||1||Included|
|E. coli RNA Feeding Strain (dpy-13) LB Amp/Tet Slant||1||Included|
|E. coli OP50 on LB Slant||1||Included|
|Wild-type Worms on NGM-lite Plates||4||Included|
|1x Lysis Buffer, 0.5 mL||1||Included|
|Sterile Bulb Pipets||6||Included|
|Sterile Culture Tubes, 15 mL||3||Included|
|Sterile Inoculating Loops||3||Included|
|LB Broth, 3 mL||3||Included|
|Mineral Oil, 5 mL||1||Included|
|Melt 'n Pour NGM-lite, 300 mL||1||Included|
|Melt 'n Pour NGM-lite, 100 mL||1||Included|
|Sterile Petri Dishes, 60 mm||40||Included|
|Ready-to-Go™ PCR Beads (0.2 mL tubes)||25||Included|
|Sterile Distilled Water, 1 mL||1||Included|
|Teacher's Manual and Reproducible Student Guide||1||Included|
|Stereomicroscope||Needed, Not included|
|Bunsen Burner||Needed, Not included|
|Containers with Cracked or Crushed Ice||Needed, Not included|
|Container with Liquid Nitrogen or Dry Ice||Needed, Not included|
|Digital or Instant Camera (optional)||Needed, Not included|
|Ethanol, 95%||Needed, Not included|
|Gel Casting Trays and Combs||Needed, Not included|
|Gel Electrophoresis Chamber||Needed, Not included|
|Masking Tape||Needed, Not included|
|Microcentrifuge||Needed, Not included|
|Microcentrifuge Adaptors||Needed, Not included|
|Micropipettor and Tips, 1-20 μL and 20-200 μL||Needed, Not included|
|Microcentrifuge Tube Rack||Needed, Not included|
|Mineral Oil||Needed, Not included|
|PCR Tubes, 0.2 or 0.5 μL||Needed, Not included|
|Permanent Markers, Black and Red||Needed, Not included|
|Power Supply||Needed, Not included|
|Steel Scalpel, Spatula, or Forceps||Needed, Not included|
|Thermal Cycler||Needed, Not included|
|UV or White Light Transilluminator||Needed, Not included|
|Water Bath, 60°C (for agarose solution)||Needed, Not included|
|Heavy-duty Plastic Bag||Needed, Not included|
|Household Bleach||Needed, Not included|
Carolina Biological and the Cold Spring Harbor Laboratory's DNA Learning Center (DNALC) have partnered together since 1986 to make molecular genetics and biotechnology more accessible to students globally. Focused on creating innovative classroom activities that give students the opportunity to go hands-on with DNA and explore the core principles of genetics. Together we have constantly worked to increase the accessibility of many key scientific breakthroughs for classrooms globally, including such techniques as PCR, DNA Sequencing, and RNAi.
Bringing a Carolina and DNALC kit into your classroom introduces your students to the experience of scientific research on its highest level, in some cases Nobel Prize winning research, with ease and simplicity. Each kit features in-depth background information, classroom tested protocols, pre-packaged materials, and the knowledgeable technical support of the Carolina Biological team. Our wide selection of kits in this series offers educators' unmatched opportunities to bring biotechnology or genetics into their classroom, with a best in class product.
At Carolina, we believe that innovative research doesn't just belong in a lab—it belongs in your classroom.
The DNA Learning Center (DNALC) is the world's first science center devoted entirely to genetics education and is an operating unit of Cold Spring Harbor Laboratory, an important center for molecular genetics research. The mission of the DNA Learning Center is to prepare students and families to thrive in the gene age. We envision a day when all elementary students are exposed to principles of genetics and disease risk; when all high school students have the opportunity to do hands-on experiments with DNA; and when all families have access to genetic information they need to make informed health care choices.
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