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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, 300¬µL), 4 mL ampicillin, proteinase K (20mg/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), 1mL sterile distilled water, 6 sterile dropping pipets, 2 melt n‚Äô 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‚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|
|Agarose, 6 g||1||Included|
|CarolinaBLU&trade Gel and Buffer Stain, 7 mL||1||Included|
|CarolinaBLU&trade Final Stain 250 mL||1||Included|
|TBE Buffer, 200 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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