I came across this technology for DNA shearing http://www.kbioscience.co.uk/instrum...ept%202008.pdf. Has anyone got any experience with this technology?
-
-
I saw this poster being presented at a recent GS-FLX user’s group meeting in London. I got the impression that 454 Branford had primarily implemented this technology in order to scale up their sample throughput. -
Using the Covaris for shearing DNA has both advantages and disadvantages.
On the plus side you can massively reduce the amount of input DNA, we’ve made shotgun libraries using 500 ng of DNA. On the down side however it’s not as reproducible as using nebulisation.
The attached are screenshots of Agilent traces (without SPRI bead clean up) where the following settings have been used:
Duty cycle = 20%
Intensity = 3
Cycle burst = 200
Time = 45 seconds.
Peak on 1 is at around 640 bp and about 100 bases lover on 2.
Both samples contain 5 ug of DNA in 100 ul of TE. The smaller sized tube was used (max capacity 130 ul) with the fiber in place. It’s also important to use the metal crimped cap as lots of the energy is lost using the plastic ones.
As things stand I wouldn't recommend switching all DNA shearing over to this system but it can be very useful when less starting material is available.Comment
-
The graphs seem to indicate that there may be a sample viscosity issue and you might want to try some of the following ideas,
-What is the mass of DNA, if it is more than 3ug it is worth trying a "pre-conditioning step"
Simply raise waterbath to 20degC and run a short "blast" 5 secs, (20% duty cycle, level 5 intensity, 200 cycles per burst) prior to the desired fragmentation dose. Up to 20ug/100ul have been run under these conditions.Last edited by Anthony; 02-26-2009, 09:15 AM.Comment
-
Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?
Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating...10-14-2026, 09:55 AM -
Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.
We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing...
Comment