Has anyone used Nextera library prep for illumina? I'm looking to test but any info on biases or other data would be welcome.
-
-
We tried it. If you don't have a Covaris it might be worthwhile. It is very easy and fast but it doesn't give a tight size distribution. -
Thanks.
We do have a covaris but it's the price. We have to charge a wet lab hourly rate. If nextera can cut 2-4 hours off library making it might be useful.
I suppose we could use an ez-gel to tighten the size range.Comment
-
I would use the HMW buffer, never the LMW buffer for Illumina prep. The peak at 200 represents 135 bp of the Nextera adapters and 65 bp of insert. I have had to use the HMW buffer and do size selection. Be careful if you do Size Selection post PCR, you do not have overages to re-amplify and the standard PE PCR Primers will not work.Comment
-
Starting with the recommended amount of DNA for Nextera (50 ng), you should get 300-500 ng of "tagmented" DNA after 9 cycles of PCR. We have some customers who don't bother with size selection, and get good sequencing results--they find that a tight size distribution is not necessary for Illumina sequencing in their application. However, it's your choice.
Also, in response to the original point, Nextera will definitely save 2-4 hours compared to Covaris + adaptor ligation.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