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Improved
methodologies for in-situ hybridization and detection of hybridized nucleic
acid sequences in cell cultures and tissue sections are provided which
offer an increase of speed, sensitivity, and simplicity unavailable in
previously known techniques. The invention detects specific nucleic acids
of interest, particularly RNA sequences, within cells and tissues utilizing
DNA of a particular size as a probe to find those sequences which are
held substantially in common between the cell or tissue and the probe.
The cells are fixed preferably in paraformaldehyde and then hybridized
using a hybridization fluid for not less than 10 minutes but not substantially
more than 24 hours. A variety of identifying labels are attached to the
probe which permit quick and rapid detection via measurement of radioactive
isotope decay or by colorimetric detection of enzymatic reaction products.
The invention is intended for use as a diagnostic kit in clinical/diagnostic
laboratory testing facilities in that it permits a relatively unskilled
person to accurately and reproducibly detect a few molecules of a specific
nucleic acid of interest in-situ in 10 minutes. |
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