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

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.