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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
  1. Caput, D., et al. (1986) Proc. Natl. Acad. Sci. USA 83: 1670-1674.

  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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