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Chimeric probe‐mediated ribonuclease protection assay for molecular diagnosis of mRNA deficiencies
Author(s) -
Maillet Philippe,
Delaunay Jean,
Baklouti Faouzi
Publication year - 1996
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1996)7:1<61::aid-humu9>3.0.co;2-s
Subject(s) - biology , cytoskeleton , ribonuclease , messenger rna , computational biology , microbiology and biotechnology , genetics , rna , gene , cell
Investigations throughout the last decade have established that cytoskeleton integrity ensures red cell deformability and mechanical stability, and that defects in one of the skeletal components usually result in more or less severe hemolytic anemias. Although a large number of molecular defects have been identified to date, many others still bypass fast and commonly used methods, such as SSCP and DGGE, mostly because of a subtle change in mRNA transcription level or a complex interaction leading to the loss of other components. We describe a ribonuclease protection assay based on a simultaneous quantification of two cytoskeletal transcripts, using a chimeric probe, emphasizing the value of a nonspecific bridging sequence, inserted between the two specific probe sequences. It is anticipated that this powerful and reliable procedure would be an additional tool in the methodology array used for screening cytoskeletal inherited abnormalities. © 1996 Wiley‐Liss, Inc.