| Title |
Gene transcript amplification from cell lysates in continuous-flow microfluidic devices
|
|---|---|
| Published in |
Biomedical Microdevices, May 2007
|
| DOI | 10.1007/s10544-007-9083-1 |
| Pubmed ID | |
| Authors |
Asensio Gonzalez, Doina Ciobanu, Michael Sayers, Noel Sirr, Tara Dalton, Mark Davies |
| Abstract |
Continuous-flow analysis, where samples circulate encapsulated in a carrier fluid is an attractive alternative to batch processing for high-throughput devices that use the polymerase chain reaction (PCR). Challenges of continuous-flow prototypes include the hydrodynamic and biological incompatibility of the carrier fluid, microchannel fouling, sample carryover and the integration of a nucleic acid extraction and reverse transcription step. We tested two homemade, continuous-flow thermocycler microdevices for amplification of reverse-transcribed messages from cell lysates without nucleic acid extraction. Amplification yield and specificity were assessed with state-of-the-art, real-time quantitative equipment. Carryover contamination between consecutive samples was absent. Amplification specificity and interference by genomic DNA were optimized by primer design. Robust detection of the low-copy transcript CLIC5 from 18 cells per microliter is demonstrated in cultured lymphoblasts. The results prove the concept that the development of micro-total analysis systems (micro-TAS) for continuous gene expression directly from cell suspensions is viable with current technology. |
Mendeley readers
The data shown below were compiled from readership statistics for 14 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Japan | 1 | 7% |
| Hungary | 1 | 7% |
| Unknown | 12 | 86% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 7 | 50% |
| Researcher | 3 | 21% |
| Student > Master | 2 | 14% |
| Lecturer | 1 | 7% |
| Student > Postgraduate | 1 | 7% |
| Other | 0 | 0% |
| Readers by discipline | Count | As % |
|---|---|---|
| Engineering | 5 | 36% |
| Agricultural and Biological Sciences | 3 | 21% |
| Biochemistry, Genetics and Molecular Biology | 2 | 14% |
| Chemistry | 2 | 14% |
| Medicine and Dentistry | 1 | 7% |
| Other | 2 | 14% |
Attention Score in Context
This research output has an Altmetric Attention Score of 3. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 14 July 2018.
All research outputs
#7,454,066
of 22,788,370 outputs
Outputs from Biomedical Microdevices
#233
of 747 outputs
Outputs of similar age
#25,290
of 72,010 outputs
Outputs of similar age from Biomedical Microdevices
#13
of 26 outputs
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