Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F17%3A00482563" target="_blank" >RIV/86652036:_____/17:00482563 - isvavai.cz</a>
Alternative codes found
RIV/61989592:15310/17:73583525
Result on the web
<a href="http://dx.doi.org/10.1093/nar/gkx588" target="_blank" >http://dx.doi.org/10.1093/nar/gkx588</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1093/nar/gkx588" target="_blank" >10.1093/nar/gkx588</a>
Alternative languages
Result language
angličtina
Original language name
Two-tailed RT-qPCR: a novel method for highly accurate miRNA quantification
Original language description
MicroRNAs are a class of small non-coding RNAs that serve as important regulators of gene expression at the posttranscriptional level. They are stable in body fluids and pose great potential to serve as biomarkers. Here, we present a highly specific, sensitive and cost-effective system to quantify miRNA expression based on two-step RT-qPCR with SYBR-green detection chemistry called Two-tailed RT-qPCR. It takes advantage of novel, target-specific primers for reverse transcription composed of two hemiprobes complementary to two different parts of the targeted miRNA, connected by a hairpin structure. The introduction of a second probe ensures high sensitivity and enables discrimination of highly homologous miRNAs irrespectively of the position of the mismatched nucleotide. Two-tailed RT-qPCR has a dynamic range of seven logs and a sensitivity sufficient to detect down to ten target miRNA molecules. It is capable to capture the full isomiR repertoire, leading to accurate representation of the complete miRNA content in a sample. The reverse transcription step can be multiplexed and the miRNA profiles measured with Two-tailed RT-qPCR show excellent correlation with the industry standard TaqMan miRNA assays (r(2) = 0.985). Moreover, Two-tailed RT-qPCR allows for rapid testing with a total analysis time of less than 2.5 hours.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10603 - Genetics and heredity (medical genetics to be 3)
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Nucleic Acids Research
ISSN
0305-1048
e-ISSN
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Volume of the periodical
45
Issue of the periodical within the volume
15
Country of publishing house
GB - UNITED KINGDOM
Number of pages
13
Pages from-to
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UT code for WoS article
000409380900007
EID of the result in the Scopus database
2-s2.0-85032874023