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.
Bibliographical noteFunding Information:
We thank Professor Juan G. Santiago for feedback and advice regarding this work. RK acknowledges support from the Jordanian-American Commission for Educational Exchange (the Binational Fulbright Commission in Jordan) through the Jordanian Visiting Post-Doctoral Scholar Fellowship. CMH acknowledges support from the National Institute of Standards and Technology (NIST) NRC Postdoctoral Associateship Program. CMH, SAM, further acknowledge support from the NIST Joint Initiative for Metrology in Biology at Stanford. The authors have declared no conflict of interest.
- Mesofluidic device
- Small RNA
- Small RNA extraction
- Small RNA quantification