Highly sensitive detection of gene expression of an intronless gene: Amplification of mRNA, but not genomic DNA by nucleic acid sequence based amplification (NASBA)
作者： Albert Heim , Stefanie Zeuke , Isabella Maria Grumbach , Bert Top
关键词: genomic DNA 、 NASBA 、 Polymerase chain reaction 、 Gene 、 Reverse transcriptase 、 Molecular biology 、 Nucleic acid 、 Genetics 、 Self-Sustained Sequence Replication 、 Biology 、 Nucleic acid amplification technique
摘要: NASBA is an isothermal nucleic acid amplification reaction that amplifies mRNA in a dsDNA background. Although similar to the sensitive reverse transcription/polymerase chain (RT-PCR) detection, not prone give false positive results caused by genomic dsDNA. Therefore, unique for detection of transcription intronless genes, which preclude strategies such as intron spanning primer pairs control RT-PCR. Using NASBA, human interferon-beta gene was demonstrated with sensitivity 10 copies, whereas 100 ng DNA gave negative result.
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