Gene transcript amplification from cell lysates in continuous-flow microfluidic devices

2007 ◽  
Vol 9 (5) ◽  
pp. 729-736 ◽  
Author(s):  
Asensio Gonzalez ◽  
Doina Ciobanu ◽  
Michael Sayers ◽  
Noel Sirr ◽  
Tara Dalton ◽  
...  
2010 ◽  
Vol 21 (41) ◽  
pp. 415603 ◽  
Author(s):  
S Gómez-de Pedro ◽  
M Puyol ◽  
J Alonso-Chamarro

Micromachines ◽  
2019 ◽  
Vol 10 (11) ◽  
pp. 744 ◽  
Author(s):  
Xiangchun Xuan

Magnetic field-induced particle manipulation is simple and economic as compared to other techniques (e.g., electric, acoustic, and optical) for lab-on-a-chip applications. However, traditional magnetic controls require the particles to be manipulated being magnetizable, which renders it necessary to magnetically label particles that are almost exclusively diamagnetic in nature. In the past decade, magnetic fluids including paramagnetic solutions and ferrofluids have been increasingly used in microfluidic devices to implement label-free manipulations of various types of particles (both synthetic and biological). We review herein the recent advances in this field with focus upon the continuous-flow particle manipulations. Specifically, we review the reported studies on the negative magnetophoresis-induced deflection, focusing, enrichment, separation, and medium exchange of diamagnetic particles in the continuous flow of magnetic fluids through microchannels.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Radhakrishna Sanka ◽  
Joshua Lippai ◽  
Dinithi Samarasekera ◽  
Sarah Nemsick ◽  
Douglas Densmore

2007 ◽  
Vol 43 (6) ◽  
pp. 2929-2931 ◽  
Author(s):  
Liviu Clime ◽  
Boris Le Drogoff ◽  
Teodor Veres

Crystals ◽  
2018 ◽  
Vol 9 (1) ◽  
pp. 12 ◽  
Author(s):  
Mathieu Gonidec ◽  
Josep Puigmartí-Luis

Materials science is a fast-evolving area that aims to uncover functional materials with ever more sophisticated properties and functions. For this to happen, new methodologies for materials synthesis, optimization, and preparation are desired. In this context, microfluidic technologies have emerged as a key enabling tool for a low-cost and fast prototyping of materials. Their ability to screen multiple reaction conditions rapidly with a small amount of reagent, together with their unique physico-chemical characteristics, have made microfluidic devices a cornerstone technology in this research field. Among the different microfluidic approaches to materials synthesis, the main contenders can be classified in two categories: continuous-flow and segmented-flow microfluidic devices. These two families of devices present very distinct characteristics, but they are often pooled together in general discussions about the field with seemingly little awareness of the major divide between them. In this perspective, we outline the parallel evolution of those two sub-fields by highlighting the key differences between both approaches, via a discussion of their main achievements. We show how continuous-flow microfluidic approaches, mimicking nature, provide very finely-tuned chemical gradients that yield highly-controlled reaction–diffusion (RD) areas, while segmented-flow microfluidic systems provide, on the contrary, very fast homogenization methods, and therefore well-defined super-saturation regimes inside arrays of micro-droplets that can be manipulated and controlled at the milliseconds scale. Those two classes of microfluidic reactors thus provide unique and complementary advantages over classical batch synthesis, with a drive towards the rational synthesis of out-of-equilibrium states for the former, and the preparation of high-quality and complex nanoparticles with narrow size distributions for the latter.


2016 ◽  
Vol 84 ◽  
pp. 63-72 ◽  
Author(s):  
Bobby Mathew ◽  
Anas Alazzam ◽  
Ghulam Destgeer ◽  
Hyung J. Sung

Author(s):  
Paul Fleming ◽  
Tara Dalton

One step reverse-transcription polymerase chain reaction (RT-PCR) assays are an attractive option for further automating gene detection assays. One-step assays can reduce hands–on-time and the risk of sample crossover and contamination. The one-step chemistries are showing increasing use in virus detection and have been reported, in some cases, to be more appropriate than their two-step counterparts [1, 2]. Previous work presented by the Stokes Institute research group outlined a micro fluidic based continuous flow instrument which performed high throughput qPCR in nanolitre sized droplets [3]. This instrument had advantages over commercially available instruments in that it could process far more than the traditional 96 or 384 reaction setup in a single run and the reaction volume was reduced from 20–50 μl down to 30–100 nl sized droplets. Combining one-step chemistry with the technology offered by the devices being developed would lead to a high-throughput RNA-to-signal system capable of reverse transcribing and performing PCR on thousands of nanolitre sized reactions every day. It is envisaged that this technology will also lead to gene expression from single cells contained in nanolitre sized droplets. In this paper, a study was conducted in which an extra thermal region, manufactured from aluminium, was added to the existing continuous flow instruments. This region was maintained at a temperature suitable for reverse transcription, which was 48°C for the one-step kit tested. The thermal region was also a suitable length to maintain the sample at the required temperature for 15 minutes. Using a commercially available one step RT-PCR kit (TaqMan® RNA-to-CT™ 1-Step Kit, 4392653), the device was evaluated for its potential to perform one-step RT-PCR in continuously flowing nanolitre sized droplets. Electrophoresis gels were initially used in assessing specific amplification before an end-point detection method was utilized. RNA was extracted from the leukemic REH cell line with the housekeeping gene, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) as the gene of interest. To investigate the possibility of further reducing sample preparation and facilitating further automation, amplification from cell lysates without nucleic acid extraction was carried out on the device. Cell lysates were prepared using the cell lysis buffer from the TaqMan® Gene Expression Cells-to-CT™ Kit (Cat #AM1728). It was found that the device was successful in one-step RT-PCR from extracted RNA samples and samples from cell lysates without nucleic acid extraction.


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