In situ transcription in cells and tissues

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 4, 4351722, 4351723, 435 91, 436 27, 935 16, 935 18, 935 19, 935 93, C12Q 168

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050213350

ABSTRACT:
In situ transcription is provided by hybridizing cells or tissue with a primer, and extending the primer bound to any template mRNA or DNA with reverse transcriptase or DNA polymerase in the presence of labeled nucleotides, which allows for detection of cells containing the template. The resulting cDNA may be eluted and used in a polymerase chain reaction for isolation of the DNA, and/or the cDNA electrophoresed, which may provide for information concerning the sequence, or the like.

REFERENCES:
patent: 4886741 (1989-12-01), Schwartz
Maniatis T., Fritch E., and Sambrook (1982), Molecular Cloning: A Laboratory Manual, pp. 213-216 and pp. 224-235.
Pardue M. L. (1987) in: Hames B & Higgins S., "Nucleic Acid Hybridisation: A Practical Approach", Chap. 8, pp. 179-202.
Eberwine, et al., "Isolation of Enzyme cDNA Clones by Enzyme Immunodetection Assay: Isolation of a Peptide Acetyltransferase", in PNAS, 84:1449-1453 (1987).
Saiki et al., "Primer-Directed Enzymatic Amplification of DNA with a Thermostable DNA Polymerase", in Science, 239:487 (1988).
Baltimore, "Viral RNA-dependent DNA Polymerase", in Nature, 226:1209 (1970).
Tecott, L. (1988) In Situ Transcription: Specific Synthesis of Complementary DNA in Fixed Tissue Sec., Science 240:1661-64.

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