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- W1576562019 abstract "Ribonucleases can specifically recognize and cleave RNA at the site of sequence mismatches in RNA-DNA or RNA-RNA hybrids. The cleavage products are then characterized by gel electrophoresis. In this unit, a procedure is presented for RNase cleavage of (32)P-labeled riboprobes (transcribed from a cloned copy of the normal sequence) that have been annealed to amplified sequences of a candidate gene or cDNA obtained from affected individuals. A Support Protocol explains how to prepare riboprobes from a genomic or cDNA template obtained from a nonmutant individual. An alternate protocol describes cleavage of RNARNA hybrids using a nonisotopic RNase cleavage mutation assay. Sequential PCR and in vitro transcription steps generate sufficient quantities of duplex RNA targets so that the cleavage products can be detected on a gel by ethidium bromide staining. The unit also discusses the use of alternative ribonucleases for cleaving singlebase mismatches." @default.
- W1576562019 created "2016-06-24" @default.
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- W1576562019 date "1997-08-01" @default.
- W1576562019 modified "2023-10-12" @default.
- W1576562019 title "Detection of Mutations by RNase Cleavage" @default.
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- W1576562019 doi "https://doi.org/10.1002/0471142905.hg0702s14" @default.
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