Keywords

blood plasma separation, qRT-PCR (quantitative Real-Time Polymerase Chain Reaction), RNA extraction, two-spin centrifuge method, hemolysis assessment

Abstract

The spinning disk technology has previously been utilized to isolate bacterial components from blood in hours instead of days. We hypothesized that this platform could be applied as an alternative approach to isolating plasma RNA from a whole blood sample. We consequently tested the efficacy of the spinning disk technology to extract plasma from whole blood upstream of RNA isolation and analysis. To do so, we collected plasma using either the spinning disk or the typical two-spin centrifuge method. We found that the spinning disk method results in significantly more hemolysis during collection than the conventional two-spin centrifuge method. However, when plasma RNA recovered from both collection methods was quantified using quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), we found that the spinning disk method yielded a higher plasma RNA concentration than the two-spin centrifuge method. This suggests that the spinning disk may be an efficient alternative method to recover plasma RNA. Further work is needed to determine whether red blood cell RNA contamination is present in the plasma RNA extracted from spinning disk-processed plasma.

Original Publication Citation

Naomi Rapier-Sharman, Mae-Lynn L. Hutchinson, Carlos Moreno, Abraham Quaye, Brian D. Poole, K. Scott Weber, William G. Pitt, and Brett E. Pickett*, “Comparison of Cell-free RNA (cfRNA) Extraction Yield from Spinning Disk vs Traditional Method”, MicroPub. Biol., Nov. 3, 2023, (2023). doi: 10.17912/micropub.biology.001007

Document Type

Peer-Reviewed Article

Publication Date

2023-11-03

Publisher

Caltech Library

Language

English

College

Ira A. Fulton College of Engineering

Department

Chemical Engineering

University Standing at Time of Publication

Full Professor

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