Droplet-based gene expression analysis using a device with magnetic force-based-droplet-handling system.
A droplet-based cell lysis and reverse transcription-polymerase chain reaction (PCR) were performed on-chip employing magnetic force-based-droplet-handling system. The actuation with a magnet offers a simple system for droplet manipulation; it does not need mechanical fluidic systems such as pumps and valves for handling solutions. It can be used as a powerful tool for various biochemical applications by moving and coalescing sample droplets using magnetic beads immersed in mineral oil. The droplet containing magnetic beads and the cells were manipulated with the magnet located underneath the channel, and coalesced with a droplet of lysis buffer. Using K562 cells as the leukemia model, the cell lysis, cDNA synthesis, and amplification of WT1 gene that is known as the prognostic factor for acute leukemia were successfully performed from a single cell.
Okochi
Mina
M
Department of Biotechnology, School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Japan.
Tsuchiya
Hiroyoshi
H
Kumazawa
Fumitaka
F
Shikida
Mitsuhiro
M
Honda
Hiroyuki
H
D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000295:instrumentation* / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D008280:Magnetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods