Isolation of enriched small RNA from cell-lysate using on-chip isotachophoresis

Ruba Khnouf, Crystal M. Han, Sarah A. Munro

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)3140-3147
Number of pages8
JournalELECTROPHORESIS
Volume40
Issue number23-24
DOIs
StatePublished - Dec 1 2019

Keywords

  • Isotachophoresis
  • Mesofluidic device
  • Small RNA
  • Small RNA extraction
  • Small RNA quantification

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't

Fingerprint Dive into the research topics of 'Isolation of enriched small RNA from cell-lysate using on-chip isotachophoresis'. Together they form a unique fingerprint.

Cite this