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Human Universal cDNA Library Array I

n and hybridization were performed in 6X SSC buffer, 5X Denhardts reagent, and 0.1% SDS. The oligonucleotide probes were prepared using T4 polynucleotide kinase and high-specific-activity [g-32P]ATP (3000 Ci/mmol). The specific activity for the probes were approximately 5 10 8 cpm/mg, and their concentrations in the hybridization solution were 5 105 cpm/ml.

The two sets of Master Membranes were hybridized overnight at 42C, and then washed in a 6X SSC buffer and 0.1% (w/v) SDS wash solution up to 1C below the calculated Tm. The membranes were mounted with Glogos II autorad markers and exposed to x-ray film with intensifying screens at 80C for 16 hours. After exposure, the autoradiograph films were realigned on the mounted membranes to locate the positive signals.

REFERENCES
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  2. Chomcynski, P. and Sacchi, N. (1987) Anal. Biochem. 172: 156.

  3. Llias, J.-M., et al. (1993) Proc. Natl. Acad. Sci. USA 90: 1479-1483.

  4. Ohmori, H. and Toyama, S. (1992) J. Cell Biol. 116 (4): 933-941.

  5. Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989) In Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.


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