Articles tagués PCR

Detecting extremely rare mutants with single-cell PCR technology

Detecting extremely rare mutants with single-cell PCR technology

Now, Richard Mathies’ group at the University of California, Berkeley (UC Berkeley) has developed a PCR-based tool for high-throughput single-cell analysis: microfabricated emulsion generator arrays (MEGAs). This device uses microfluidic technology to undertake massively parallel PCR amplification of DNA from single cells, providing accurate detection and quantification of rare mutants within a population (1). The use of small water droplets in oil to amplify single molecules is known as emulsion PCR; its power comes from the ability to miniaturize reactions into droplets and from using primer-coupled beads (2,3). In this way, all PCR products are the same (or clonal) and physically attached to the bead for easy retrieval.


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Palm PCR

[update] Vous pouvez le voir sur aminopop, en quatre couleurs ! Ouah!, m’en faudrait un bleu siouplait
Puis je me dis que les faucheurs volontaires pourraient s’en servir pour identifier les champs de plates trasgéniques, pour ne pas risquer des victimes innocentes de toute contamination 😀 [Update]


L’arrivée des portables et des mobiles (ordinateurs et téléphones) a fait que le bureau a pu nous suivre durant les périodes des vacances. Au moins aux endroits où la couverture du sans fil et éventuellement de la WiFi est suffisante. J’en connais qui poussés par leur envie de rester connectés se trimbalaient entre autres dans le coffre de la voiture un générateur pour charger leur portable, le temps que les chargeurs à travers l’allume-cigare soient performants. Et d’autres qui faisaient en sorte que leur mobile soit éteint durant toute la période des vacances, pour pouvoir profiter des pdf qu’ils avaient embarqué dans leur portable.

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The Tables serie: Table 1-Highly Integrated Molecular Diagnostic Platforms

This is the 1st table of a serie of 3 on automated platforms in Molecular Diagnostics that will be available exclusively at Coffee And Sci(ence).  You can find it here:
Table 1-Highly Integrated Molecular Diagnostic Platforms 
Back in 2004, when Roche Molecular Diagnostics had introduced the Cobas Ampliprep + Cobas Taqman 48 in Europe, there was not too much instruments capable of such integration of nuclec acid extraction (NA) and realtime PCR (rtPCR) detection. Now (Nov. 2007), there is a bunch of them. Abbott, BD, Biomerieux, Cepheid, Gen-Probe, HandyLab, Iquum, Qiagen, Roche Diagnostics and Siemens Medical Solutions Diagnostics. If you are not here, please contact us.
Integration is essentially coming from the combination of 2 instruments, 1 for NA extraction, one for rtPCR. Abbott, Biomerieux, Roche and Siemens are the majors. They basically use Tecan, Hamilton, PSS based platforms for NA extraction combined with a rtPCR thermocycler from AB, former Organon-Teknika, Roche, Stratagene. Transfer of sample between instruments is manual, except for Roche who has produced the Docking Station, a robotic interface whichacts as a PCR plate conveyor.
I have mentionned Qiagen in this table because they have so much NA extraction platforms available and a long list of Molecular Diagnostic (MDX) reagents. However, Qiagen is still missing a rtPCR platform. Artus Biotech, before their acquisition by Qiagen, had a collaboration with Corbett Research around the RotorGene 3000. But Qiagen is not mentionning this partnership anymore. Hopefully they are on the market to find a solution.
Gen-Probe and BD have a different approach in that sense they are fully integrated instruments in a single box. Gen-Probe’s Tigris is the precursor of such category of instruments. Respect for that. But come on, this is a « Qual » platform, not a Quant. BD’s Viper is almost of a same age, however, BD has added very recently this fully integrated capacity to the Viper which is now capable of sample prep, SDA setup, and detection. But the specs of this later are still unclear. There is no mention at all of this platform in the real world, except in meetings and on BD’s booth. Is that rtSDA? I’m not sure.
Finally, a more recent category of platforms, the benchtop fully integrated MDX platforms, with Cepheid, Iquum and HandyLab. Cepheid is here the precursor with the GeneXpert4. They recently launched the GX1 and GX16 which are the younger and bigger bros of the GX4. Iquum is working on its Lab-in-a-tube technology since 2004. Hopefully we’ll see the Liat available soon. HandyLab has shown more recently their PCR chip based technology within their upcoming Jaguar platform.
Whoever will succeed the best, Cepheid, HandyLab and Iquum are clearly building the future of MDX platforms. Nanosphere’s Verigene system who has recently been approved by the FDA for various MDX assays, may also be part of the game. See this blog elsewhere, and remember Nanosphere is not using anymore PCR steps. 

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