RT-Hpro-PCR: A MicroRNA Detection System Using a Primer with a DNA Tag

被引：10

作者：

Takei, Fumie ^{[1
]}

Akiyama, Misaki ^{[2
]}

Murata, Asako ^{[2
]}

Sugai, Ayako ^{[2
]}

Nakatani, Kazuhiko ^{[2
]}

Yamashita, Ichiro ^{[3
]}

机构：

[1] Natl Def Med Coll, 3-2 Namiki, Tokorozawa, Saitama 3598513, Japan

[2] Osaka Univ, Inst Sci & Ind Res, 8-1 Mihogaoka, Ibaraki, Osaka 5670047, Japan

[3] Osaka Univ, Grad Sch Engn, 8-1 Mihogaoka, Ibaraki, Osaka 5670047, Japan

来源：

CHEMBIOCHEM | 2020年 / 21卷 / 04期

基金：

日本科学技术振兴机构;

关键词：

C-bulge DNA; fluorescence; hairpin probes; microRNAs; RNA recognition; RT-PCR; BULGE; DYSREGULATION; CHEMISTRY; PROBES;

D O I：

10.1002/cbic.201900382

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：477 / 480

页数：4

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