Isolation of enriched small RNA from cell-lysate using on-chip isotachophoresis.
Electrophoresis
; 40(23-24): 3140-3147, 2019 12.
Article
en 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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Técnicas Analíticas Microfluídicas
/
ARN Pequeño no Traducido
/
Isotacoforesis
Límite:
Humans
Idioma:
En
Revista:
Electrophoresis
Año:
2019
Tipo del documento:
Article
País de afiliación:
Estados Unidos