EZ-Bead system

[rtpon]

Has anyone used the EZ-Bead system? Does it work well? We are considering purchase of either an Illumina HiSeq or ABI SOLiD 4hq and while both instruments have their merits, the ABI bead preparation seems a lot more laborious. Does the EZ-Bead system really help simplify the work?

[FLeader]

We recently obtained one and it seems to work as well as advertised. It now takes a couple of hours of hands-on time as compared to a couple of days to go from library to slides. Bead quality seems excellent. We will have to run a few more samples to see if any issues crop up, but so far we are very happy.

[dsidote]

We have one too and our experiences are the same as FLeader. We have run 6 samples so far and all have been excellent.

[bioits]

Comparing to the manual prep, EZ bead is so much easier and works well so far.

[VanessaS]

I love love LOVE LOVE LOVE my EZBead systems. LOVE. The hands on time is so minimal, which not only frees you up to do more library prep, etc, but it also minimizes human error. My bead quality is fantastic and consistant, I really couldn't ask for more. Oh, wait, yes I could. I want more EZBead systems please!

[pmiguel]

We don't have an EZBead system yet. But while it will likely drastically reduce the effort needed to create and enrich templated beads, tailing and bead deposition will still be necessary. This is extra work for which I do not believe there is an equivalent in the HiSeq workflow. However, though extra effort, it might be beneficial in some circumstances as it might represent a flexibility not present in the HiSeq.

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Phillip

[rtpon]

Thank you everyone for your advice.

[fbettoni]

enrichment percentage

I'm trying to calculate the enrichment percentage with the EZbead system. Does anybody know how I can do this?

[VanessaS]

I think what you are looking for is on the WFA, the p2#/p1# ratio.

[pmiguel]

We count the enriched and the non-enriched beads to get the % enrichment on the EZ Bead.

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Phillip

[fbettoni]

Ok but before the WFA how do you determine the number of beads you have? First, what is the inicial amount of beads used (E80)? When considering the enrichment, where do you take the beads from (which bottle) and what is the volume before and after the enrichment?
Can I count using Nanodrop or do I have to use FACS?

[pmiguel]

We use the nanodrop to "count" beads. Determine the number of beads/ul and multiply by the total number of ul.

You can recover the non-enriched beads from the container they are decanted into by the EZBead. Or you can take an aliquot of the beads prior to ePCR and determine the total number of beads that are going into ePCR initially.

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Phillip