Result: An integrated microfluidic system for automated extraction of in vitro transcribed mRNAs via probe-coated magnetic beads.

Title:
An integrated microfluidic system for automated extraction of in vitro transcribed mRNAs via probe-coated magnetic beads.
Authors:
Gurme ST; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. gwobin@pme.nthu.edu.tw., Su YT; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. gwobin@pme.nthu.edu.tw., Koparde B; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. gwobin@pme.nthu.edu.tw., Wang LH; School of Medicine, National Tsing Hua University, Hsinchu, Taiwan. wang.huiching@gmail.com., Lee GB; Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan. gwobin@pme.nthu.edu.tw.; Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu, Taiwan.
Source:
Lab on a chip [Lab Chip] 2026 Jan 20; Vol. 26 (2), pp. 471-480. Date of Electronic Publication: 2026 Jan 20.
Publication Type:
Journal Article
Language:
English
Journal Info:
Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 101128948 Publication Model: Electronic Cited Medium: Internet ISSN: 1473-0189 (Electronic) Linking ISSN: 14730189 NLM ISO Abbreviation: Lab Chip Subsets: MEDLINE
Imprint Name(s):
Original Publication: Cambridge, UK : Royal Society of Chemistry, c2001-
MeSH Terms:
RNA, Messenger*/isolation & purification , RNA, Messenger*/genetics , Lab-On-A-Chip Devices* , Microfluidic Analytical Techniques*/instrumentation , Microfluidic Analytical Techniques*/methods , Microspheres* , Transcription, Genetic*, Humans ; Automation
Substance Nomenclature:
0 (RNA, Messenger)
Entry Date(s):
Date Created: 20251223 Date Completed: 20260120 Latest Revision: 20260120
Update Code:
20260121
DOI:
10.1039/d5lc00938c
PMID:
41431383
Database:
MEDLINE

Further Information

Extraction and purification of mRNA are critical steps for diverse molecular biological applications, including gene expression analyses, vaccine development, and genomic assays. To enhance mRNA extraction efficiency, we designed an integrated microfluidic chip capable of isolating in vitro transcribed (IVT) mRNAs (up to 93%) via probe-coated magnetic beads without the need for high-speed centrifugation, organic solvents, or cooling conditions. mRNA-specific probe-coated magnetic beads were employed, thereby automating the entire workflow, including IVT, DNase treatment, mRNA extraction, and reverse transcription (RT) could all be performed on-chip. This advancement is particularly significant for on-chip transcription-translation coupled with puromycin linker association (TRAP) display screening applications. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis confirmed the device's ability to selectively capture mRNA with a high capture efficiency of 93% within 15 minutes and 48% of thermal release efficiency after 10 minutes at 95 °C. By incorporating this optimized approach into a fully automated lab-on-chip system, puromycin-linked mRNAs could be directly used for peptide screening of kinesin family member 2C (KIF2C), a protein overexpressed in aggressive cancers.