TY - JOUR
T1 - Ultrasensitive detection of miRNA via one-step rolling circle-quantitative PCR (RC-qPCR)
AU - Xu, Mingcheng
AU - Ye, Jiawei
AU - Yang, Dan
AU - Abdullah AL-maskri, Abdu Ahmed
AU - Hu, Haihong
AU - Jung, Cheulhee
AU - Cai, S.
AU - Zeng, Su
N1 - Funding Information:
We acknowledge financial support from the National Natural Science Foundation of China (Grant 81573389 ), the National Key R&D Program of China ( 2017YFC0908600 ), the Zhejiang Provincial Natural Science Foundation of China ( LY18H300003 ). Scientific Research Fund of Zhejiang Provincial Education Department ( Y201430444 ).
PY - 2019/10/24
Y1 - 2019/10/24
N2 - A novel microRNA (miRNA) quantification method has been developed using one-step rolling circle-quantitative PCR (RC-qPCR) analysis. Vent (exo-) DNA polymerase is firstly utilized to combine a rolling circle amplification (RCA) and qPCR in one step with high sensitivity and specificity in our RC-qPCR assay. Before performing the RC-qPCR, a padlock probe is ligated only when it is perfectly hybridized with miRNA. This ligation-based miRNA assay is highly specific for mature miRNAs, discriminating among related miRNAs that differ by as little as one nucleotide. It exhibits a dynamic range of seven orders of magnitude with a detection limit of 500 aM, and could be also used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs).
AB - A novel microRNA (miRNA) quantification method has been developed using one-step rolling circle-quantitative PCR (RC-qPCR) analysis. Vent (exo-) DNA polymerase is firstly utilized to combine a rolling circle amplification (RCA) and qPCR in one step with high sensitivity and specificity in our RC-qPCR assay. Before performing the RC-qPCR, a padlock probe is ligated only when it is perfectly hybridized with miRNA. This ligation-based miRNA assay is highly specific for mature miRNAs, discriminating among related miRNAs that differ by as little as one nucleotide. It exhibits a dynamic range of seven orders of magnitude with a detection limit of 500 aM, and could be also used for the quantification of other small RNA molecules such as short interfering RNAs (siRNAs).
KW - MiR-200a
KW - MiRNA detection
KW - One-step amplification
KW - Rolling circle amplification
KW - Rolling circle-quantitative PCR (RC-qPCR)
KW - Vent (exo-) DNA polymerase
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U2 - 10.1016/j.aca.2019.05.028
DO - 10.1016/j.aca.2019.05.028
M3 - Article
C2 - 31307711
AN - SCOPUS:85065848277
VL - 1077
SP - 208
EP - 215
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
SN - 0003-2670
ER -