期刊
CHEMBIOCHEM
卷 21, 期 4, 页码 477-480出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201900382
关键词
C-bulge DNA; fluorescence; hairpin probes; microRNAs; RNA recognition; RT-PCR
资金
- JSPS [16K14 027]
- Center of Innovation Program from the Japan Science and Technology Agency (JST)
- Five-star Alliance in NJRC Mater. Dev. [20183056]
MicroRNAs (miRNAs) are short RNAs that regulate the expression of complementary messenger RNAs and are involved in numerous human diseases. However, current detection techniques lack the sensitivity to detect miRNAs of low abundance. Moreover, at a length of 20-25 bases, miRNAs are too short for the reverse transcription (RT) polymerase chain reaction (PCR). Here we have developed a new, rapid, and simple miRNA detection system utilizing an RT primer containing a DNA tag at the 5 '-end to increase the length of the cDNA. This strategy increases the length of the hybridized tagged primer and the complementary template DNA, as well as the melting temperature of the primer.template DNA duplex. PCR efficiency is thus increased, thereby enhancing miRNA detection sensitivity.
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