TY - JOUR
T1 - RT-Hpro-PCR
T2 - A MicroRNA Detection System Using a Primer with a DNA Tag
AU - Takei, Fumie
AU - Akiyama, Misaki
AU - Murata, Asako
AU - Sugai, Ayako
AU - Nakatani, Kazuhiko
AU - Yamashita, Ichiro
N1 - Funding Information:
This work was supported by a Grant in Aid for Scientific Research Challenging Exploratory Research from the JSPS (16K14 027) to F.T., The Five-star Alliance in NJRC Mater. & Dev. (20183056) to F.T., and The Center of Innovation Program from the Japan Science and Technology Agency (JST) to I.Y.
Publisher Copyright:
© 2019 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2020/2/17
Y1 - 2020/2/17
N2 - 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.
AB - 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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U2 - 10.1002/cbic.201900382
DO - 10.1002/cbic.201900382
M3 - Article
C2 - 31397042
AN - SCOPUS:85074818919
VL - 21
SP - 477
EP - 480
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 4
ER -
