Open AccessHighly sensitive detection of gene expression of an intronless gene: amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)
Author(s) -
Albert Heim,
Stefanie Zeuke,
Isabella M. Grumbach,
Bert Top
Publication year - 1998
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/26.9.2250
Subject(s) - nasba , biology , genomic dna , microbiology and biotechnology , gene , primer (cosmetics) , polymerase chain reaction , intron , dna , nucleic acid , messenger rna , transcription (linguistics) , multiple displacement amplification , loop mediated isothermal amplification , dna microarray , gene expression , nucleic acid thermodynamics , reverse transcriptase , complementary dna , nucleic acid sequence , genetics , rna , linguistics , chemistry , philosophy , organic chemistry , dna extraction
NASBA is an isothermal nucleic acid amplification reaction that amplifies mRNA in a dsDNA background. Although similar to the sensitive reverse transcription/polymerase chain reaction (RT-PCR) in mRNA detection, NASBA is not prone to give false positive results caused by genomic dsDNA. Therefore, NASBA is unique for sensitive detection of transcription of intronless genes, which preclude strategies such as intron spanning primer pairs to control false positive results in RT-PCR. Using NASBA, mRNA of the intronless human interferon-beta gene was demonstrated with a sensitivity of 10 copies, whereas 100 ng genomic DNA gave a negative result.
Accelerating Research
Robert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom
Address
John Eccles HouseRobert Robinson Avenue,
Oxford Science Park, Oxford
OX4 4GP, United Kingdom