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MORIMASA MATSUTA AND MAYUMI MATS UTA
- A bench-top micro centrifuge
- Microcentrifuge tubes: 0.5 ml, 1 ml
- Diamond tipped pen
- Glass slides: aminoalkylsilane-coated, commercially available.
- Glass cover slips; 18X18 mm, 22X40~60 mm
- Plastic cover slips; 22X40~60 mm, cut from polyethylene bag
- Coplin jars
- Micropipettes (variable type:0.5~10111, ~200111, 1ml)
- Moist chamber
- Incubator at 37°C
- Hot plate for Rapid Hybridization and Co denaturation procedures
- Chromosome-specific, locus-specific probes; also commercially available from Vysis (Downers Grove, IL), Oncor
(Geithersburg, MD), etc.
- Master Mix (of Hybridization): Master Mix gives a hybridization mixture that is 55% formamidellO% dextran sulfate/
IXSSC.
- Purified water
- Formamide: Ultra pure grade (Kodak etc.)
- Rubber cement: Paper bond™ (Kokuyo, Tokyo, Japan)
- Denaturation solution: 50 ml of 70% formamide/2X SSC
- Washing solution: 150 ml of 50% fromamide/2X sse
- Washing solution for rapid hybridization protocol: O.4X SSC
- 20X SSC solution
- PN buffer
- PNM buffer: 5% nonfat dry milk to PN buffer + 0.02% NaAzide.
- Antifade solution (see Recipes, also commercially available)
- 2'-6-diamino-2-phenyl-indole (DAPI): 100 ng (- 1000 ng for
tissue sections) in antifade solution; this is required when
either green-fluorochrome (FITC, Spectrum Green, etc.)labelled probes or red-fluorochrome (TexasRed, Sprectrum
Orange, etc. )- labelled probes are used as counterstains
- Propidium iodide (PI): 5 - 10 llg in antifade solution, counterstain for green-fluorochrome (FITC, Spectrum Green, etc.) -
labelled probes.
- Ethanol dehydrate solution: concentrations of 70, 85 and
100%.
MORIMASA MATSUTA AND MAYUMI MATS UTA
- A bench-top micro centrifuge
- Microcentrifuge tubes: 0.5 ml, 1 ml
- Diamond tipped pen
- Glass slides: aminoalkylsilane-coated, commercially available.
- Glass cover slips; 18X18 mm, 22X40~60 mm
- Plastic cover slips; 22X40~60 mm, cut from polyethylene bag
- Coplin jars
- Micropipettes (variable type:0.5~10111, ~200111, 1ml)
- Moist chamber
- Incubator at 37°C
- Hot plate for Rapid Hybridization and Co denaturation procedures
- Chromosome-specific, locus-specific probes; also commercially available from Vysis (Downers Grove, IL), Oncor
(Geithersburg, MD), etc.
- Master Mix (of Hybridization): Master Mix gives a hybridization mixture that is 55% formamidellO% dextran sulfate/
IXSSC.
- Purified water
- Formamide: Ultra pure grade (Kodak etc.)
- Rubber cement: Paper bond™ (Kokuyo, Tokyo, Japan)
- Denaturation solution: 50 ml of 70% formamide/2X SSC
- Washing solution: 150 ml of 50% fromamide/2X sse
- Washing solution for rapid hybridization protocol: O.4X SSC
- 20X SSC solution
- PN buffer
- PNM buffer: 5% nonfat dry milk to PN buffer + 0.02% NaAzide.
- Antifade solution (see Recipes, also commercially available)
- 2'-6-diamino-2-phenyl-indole (DAPI): 100 ng (- 1000 ng for
tissue sections) in antifade solution; this is required when
either green-fluorochrome (FITC, Spectrum Green, etc.)labelled probes or red-fluorochrome (TexasRed, Sprectrum
Orange, etc. )- labelled probes are used as counterstains
- Propidium iodide (PI): 5 - 10 llg in antifade solution, counterstain for green-fluorochrome (FITC, Spectrum Green, etc.) -
labelled probes.
- Ethanol dehydrate solution: concentrations of 70, 85 and
100%.