29 Spectral Imaging Analysis of Metaphase and Interphase Cells
365
- Dry bath (heat block) (Model 2001, Labline Instruments)
- Fluorescence microscope (Nikon E600, Nikon) equipped with 40x and
60x oil immersion lenses
- Incubator oven (FISHER, set to 37°C)
- Phase microscope (Nikon E800, Nikon)
- Pulsed Field Gel Electrophoresis (PFGE) system (BIO-RAD)
- SD200 SpectraCube spectral imaging system (Applied Spectral Imaging, Inc.)
- Shaking Incubators (New Brunswick): 30°C for yeast cell culture, 37°C
for the culture of E. coli
- Thermal cycler for in vitro DNA amplification (model 4800, Perkin
Elmer)
- Mini-Fluorometer (TKO 100, Pharmacia)
- Water bath (VWR)
- ~-Agarase and buffer (New England Biolabs). Store at -20°e.
Chemicals
- 4 dNTP mix: dATP, dCTP, dGTP, and dTTP. Make 10 mM of each
dNTP solution by combining 10 I.d of a 100 mM solution (Roche Molecular Biochemical) with 60 J.lI of double distilled water. Store in 100 J.lI
aliquots at -20°e.
- Agarose (Life Technologies).
- Bovine serum albumin (Sigma). Store at 4°e.
- Co1cemid (Life Technologies), 10 mg/ml. Store at 4°C.
- Cy3-dUTP (Amersham), 1 mM. Store at -20°C.
- Digoxigenin-ll-dUTP (Roche Molecular Biochemical), 1 mM. Store at
-20°e.
- EDTA (ethylene diamine tetraacetic acid) (Life Technologies), 0.5 M,
pH 8.0.
- Ethidium bromide (Life Technologies), 10 mg/ml.
- Fetal bovine serum (Sigma). Store at -20°e.