Jump to main content
Jump to site search
PLANNED MAINTENANCE Close the message box

Scheduled maintenance work on Wednesday 27th March 2019 from 11:00 AM to 1:00 PM (GMT).

During this time our website performance may be temporarily affected. We apologise for any inconvenience this might cause and thank you for your patience.

Issue 41, 2014
Previous Article Next Article

Accelerated DNA recombination on a functionalized microfluidic chip

Author affiliations


Genetic recombination is a powerful tool to create functional DNA hybrids with high controllability and precision for applications such as target gene therapy and molecular diagnostics, but it is often limited by the slow and low-efficient reconstruction of novel DNA species, especially in bulky volume solutions. Here, we present a microfluidic platform to enable rapid and consecutive on-line DNA digestion and ligation. The microdevice, embedded with a temperature sensor, offers controllable temperature and confined space to enhance intermolecular collision, and thus improves the slicing and splicing of DNA strands of interest. The stepwise functionalization of the microchannel surface with APTES, biotin and streptavidin permits the immobilization of biotinylated double stranded DNA (dsDNA) labeled with fluorescent probes at the ends for real-time monitoring. Digestion enzyme (BamH1) and DNA T4 ligase, subsequently, flow through the channel to hydrolyze and ligate the target DNA segments separately, and both digestion and ligation occur to a large extent within 10 min, with an efficiency of up to 71% (maximal at 1 h) and 63% (maximal at 2 h) of the highest value, respectively. Compared to conventional molecular diagnostics methods, which take several hours for digestion and ligation and deplete large volumes of reagents, such a system requires minimal time and sample volumes that are desirable in low-cost molecular therapy and point-of-care tests.

Graphical abstract: Accelerated DNA recombination on a functionalized microfluidic chip

Back to tab navigation

Publication details

The article was received on 10 Mar 2014, accepted on 25 Apr 2014 and first published on 25 Apr 2014

Article type: Paper
DOI: 10.1039/C4RA02076F
Citation: RSC Adv., 2014,4, 21541-21547

  •   Request permissions

    Accelerated DNA recombination on a functionalized microfluidic chip

    F. Yang, Y. Zhang, S. Rafeah, H. Ji, S. Xie, Y. Ning and G. Zhang, RSC Adv., 2014, 4, 21541
    DOI: 10.1039/C4RA02076F

Search articles by author