Isolation of enriched small RNA from cell-lysate using on-chip isotachophoresis.
Electrophoresis
; 40(23-24): 3140-3147, 2019 12.
Article
em En
| MEDLINE
| ID: mdl-31675123
ABSTRACT
In spite of the growing interest in the roles and applications of small RNAs (sRNAs), sRNA isolation methods are inconsistent, tedious, and dependent on the starting number of cells. In this work, we employ ITP to isolate sRNAs from the cell-lysate of K562 (chronic myelogenous leukemia) cells in a polydimethylsiloxane (PDMS) mesofluidic device. Our method specifically purifies sRNA of <60 nucleotides from lysate of a wide range of cell number spanning from 100 to 1 000 000 cells. We measured the amount of sRNA using the Agilent Bioanalyzer and further verified the extraction efficiency by reverse transcription quantitative PCR. Our method was shown to be more efficient in sRNA extraction than commercial sRNA isolation kits, especially when using smaller numbers of starting cells. Our assay presents a simple and rapid sRNA extraction method with 20 min assay time and no intermediate transfer steps.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Técnicas Analíticas Microfluídicas
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Pequeno RNA não Traduzido
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Isotacoforese
Limite:
Humans
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article