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BIOLUMINESCENT AMPLIFICATION ASSAY FOR microRNA DETECTION
Author(s) -
Sun Ye,
Golovlev Val
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.914.3
Subject(s) - microrna , multiple displacement amplification , rolling circle replication , loop mediated isothermal amplification , dna microarray , microbiology and biotechnology , dna , detection limit , bioluminescence , chemistry , biology , polymerase chain reaction , chromatography , dna replication , biochemistry , gene , dna extraction , gene expression
We have previously reported a new sensitive and cost effective Reverse Transcription Quantitative Bioluminescent assay (RT‐qBLA) for detection and quantification of microRNA in biological samples 1 , 2 . The RT‐qBLA detects release of inorganic pyrophosphate when target microRNA is extended on specially designed DNA template. The proposed approach is fast and less expensive than microarrays and real‐time PCR for microRNA detection. It allows microRNA quantification in a broad dynamic range; requires less reagents per reaction; can be performed at a constant temperature and is amendable to various replication strategies including “constant temperature” amplification techniques such as Rolling Circle Amplification (RCA) and Strand Displacement Amplification (SDA). In this report we present development of a real‐time RCA and SDA for microRNA detection with enhanced detection sensitivity and specificity.