Method and assay for detection of the expression of allele-speci

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

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435 405, 435 912, C12Q 168, C12P 1934

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058043837

ABSTRACT:
A method and an allele-specific in situ reverse transcriptase polymerase chain reaction (RT-PCR) amplification assay for detection and differentiation between expression of unmutated wild-type DNA sequences and between endogenous mutated DNA sequences in vitro or in vivo. The method is useful for verification, diagnostic assessment and monitoring of therapeutic small fragment homologous replacement gene therapy, for diagnostic assessment and monitoring of cDNA-based gene therapies, for analysis of gene expression of specific alleles during fetal development and for diagnostic assessment of the expression of alleles involved in cancer mutations.

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Alfredo Martinez, Mae-Jean Miller, Kathryn Quinn, kEdward J. Unsworth, Masahito Ebina and Frank Cuttitta, Non-radioactive Localization of Nucleic Acids by Direct In Situ PCR and In Situ RT-PCR in Paraffin-embedded Sections, The Histochem. Cytochem., 43: (8):739-746, (1995).
I. A. Teo and S. Shaunak, Polymerase chain reaction in situ: an appraisal of an emerging technique, Biochemical Journal, 27:647-659, (1995).
Gerard Nuovo, In Situ RT-PCR Detection of Gelatinase mRNA in HeLa Cells on the GeneAmp In Situ PCR System 1000, Research News, 1-5, (Mar. 1995).
Gerard Nuovo, M.D., RT In Situ with Direct Incorporation of Digoxigenin-11-dUTP: Protocol and Applications, Biochemica, 11, 1:4-61, (Winter 1994).
Staecker, M. Cammer, R. Rubinstein, T.R. Van De Water, A Procedure for RT-PCR Amplification of mRNAs on Histological Specimens, BioTechniques, 16:76-80 (1994).
Rosenfeld et al. (1992) In vivo transfer of the human cystic fibrosis transmembrane conductance regulator gene to the airway epithelium. Cell 68:143-155, Jan. 1992.

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