+ Site Statistics
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Automatic extraction and processing of small RNAs on a multi-well/multi-channel (M&M) chip

Automatic extraction and processing of small RNAs on a multi-well/multi-channel (M&M) chip

Analyst 137(23): 5546-5552

The study of the regulatory roles in small RNAs can be accelerated by techniques that permit simple, low-cost, and rapid extraction of small RNAs from a small number of cells. In order to ensure highly specific and sensitive detection, the extracted RNAs should be free of the background nucleic acids and present stably in a small volume. To meet these criteria, we designed a multi-well/multi-channel (M&M) chip to carry out automatic and selective isolation of small RNAs via solid-phase extraction (SPE), followed by reverse-transcription (RT) to convert them to the more stable cDNAs in a final volume of 2 μL. Droplets containing buffers for RNA binding, washing, and elution were trapped in microwells, which were connected by one channel, and suspended in mineral oil. The silica magnetic particles (SMPs) for SPE were moved along the channel from well to well, i.e. in between droplets, by a fixed magnet and a translation stage, allowing the nucleic acid fragments to bind to the SMPs, be washed, and then be eluted for RT reaction within 15 minutes. RNAs shorter than 63 nt were selectively enriched from cell lysates, with recovery comparable to that of a commercial kit. Physical separation of the droplets on our M&M chip allowed the usage of multiple channels for parallel processing of multiple samples. It also permitted smooth integration with on-chip RT-PCR, which simultaneously detected the target microRNA, mir-191, expressed in fewer than 10 cancer cells. Our results have demonstrated that the M&M chip device is a valuable and cost-saving platform for studying small RNA expression patterns in a limited number of cells with reasonable sample throughput.

(PDF emailed within 0-6 h: $19.90)

Accession: 051728135

Download citation: RISBibTeXText

PMID: 23001054

DOI: 10.1039/c2an35883b

Related references

Automatic sleep staging using multi-dimensional feature extraction and multi-kernel fuzzy support vector machine. Journal of Healthcare Engineering 5(4): 505-520, 2015

Robust, globally consistent and fully automatic multi-image registration and montage synthesis for 3-D multi-channel images. Journal of Microscopy 243(2): 154-171, 2011

Improved multi-channel multi-carrier generation using gain-independent multi-channel frequency shifting recirculating loop. Optics Express 20(28): 29599-29604, 2013

Software development of multi-element transient signal acquisition and processing with multi-channel ICP-AES. Guang Pu Xue Yu Guang Pu Fen Xi 20(1): 55-57, 2003

Multi-thread application for multi-channel signal processing and interpretation for laser Doppler flowmetry. Biomedizinische Technik. Biomedical Engineering 43 Suppl: 102-103, 1998

Multi-channel multi-carrier generation using multi-wavelength frequency shifting recirculating loop. Optics Express 20(20): 21833-9, 2013

Overview of the Alaskan experience with multi-scale, multi-project, multi-sensor real-time geophysical data processing. Seismological Research Letters 71(2): 233, 2000

Performance evaluation of a multi-purpose mobile starch extraction plant for small scale processing of tuber crops. Journal of Root Crops 31(2): 106-110, 2005

Multi-domain feature extraction for small event-related potentials through nonnegative multi-way array decomposition from low dense array EEG. International Journal of Neural Systems 23(2): 1350006-1350006, 2013

A wirelessly programmable chip for multi-channel neural stimulation. Conference Proceedings 2012: 6595-6599, 2013

The multi-organ chip--a microfluidic platform for long-term multi-tissue coculture. Journal of Visualized Experiments 98): E52526-E52526, 2016

Analysis of multi-channel data in the peripheral nervous system--multi-channel transducer characteristics of mechanoreceptors in the frog's sartorius muscle. Iyo Denshi to Seitai Kogaku. Japanese Journal of Medical Electronics and Biological Engineering 17(1): 45-52, 1979

Automated glacier surface velocity using multi-image/multi-chip MIMC feature tracking. Eos, Transactions, American Geophysical Union 90.52, Suppl, 2009

The comparison of dispersive solid phase extraction and multi-plug filtration cleanup method based on multi-walled carbon nanotubes for pesticides multi-residue analysis by liquid chromatography tandem mass spectrometry. Journal of Chromatography. A 1385: 1-11, 2015

Cross-talk problem on a fluorescence multi-channel microfluidic chip system. Biomedical Microdevices 7(3): 205-211, 2005