[BioDynami]

SPRI Beads (DNA & RNA Purification)
Optimized to purify genomic DNA, DNA fragments >100 bp, and RNA fragments >200 bases.

Features:

 Cost effective alternative to AMPure XP and SPRIselect

• Save 75% of the cost

 Purification of DNA and RNA

• Genomic DNA and DNA fragments > 100 bp
• RNA fragments > 200 nt

Learn More

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[itsmisterbrown]

BC has released some data and recommendations in their user manual for XP beads, see FAQs# 5 and 6

https://www.beckmancoulter.com/wsrportal/techdocs?docname=B37419

[snetmcom]

Ah. So the genomic DNA stays attached to the beads, but still works fine for downstream manipulation. Interesting...

--
Phillip

This does work, but the beads will inhibit some reactions.

[pmiguel]

Ah. So the genomic DNA stays attached to the beads, but still works fine for downstream manipulation. Interesting...

--
Phillip

[melop]

Hi melop,
What are your elution conditions?
--
Phillip

I elute in 30ul of Tris pH=8.0. I pipette it until the beads get re-suspended, and directly used that mixture for down-stream workflows without removing the beads.

If you want to remove the beads, incubate at 37C overnight.

[pmiguel]

Hi melop,
What are your elution conditions?
--
Phillip

[melop]

I know this thread is old but I'm hoping Melop could tell me the ratio of sample to beads used for genomic DNA purification. I looked around online and since the protocols aren't designed for DNA this size I haven't found any suggestions.
Thanks.

You can do as low as 0.6X beads. I usually use 1X. It usually doesn't matter too much for genomic extracts. I also keep the beads in, they don't affect with PCR or library prep.

[urchin]

genomic with spri beads

I know this thread is old but I'm hoping Melop could tell me the ratio of sample to beads used for genomic DNA purification. I looked around online and since the protocols aren't designed for DNA this size I haven't found any suggestions.
Thanks.

[melop]

I'll echo ECO and melop in saying that I've tried SPRI beads (Ampure and Seramag speedbeads) to clean up WGA products as well as genomic DNA from home-brew library prep and genomic DNA extraction protocols. As ECO mentioned, they do tend to clump and stick together with higher molecular weight fragments. It might help slightly if you avoid drying the bead pellet till it cracks. Longer elution times (I often incubate overnight at 4 Celsius for genomics extracts) and a small amount of Tween-20 seems to help elute more of the bound DNA.

Yes - I agree. Didn't add tween 20 so recovery is probably lower than optimal, but it's more than enough for what I need so can't complain. Attached is a gel picture of genomic DNA extracted directly after SDS + proteinase K digestion without phenol-chloroform wash from a small piece of fish finclip (1mm x 4 mm). RNA is co-purified because I didn't treat with RNase. Qubit shows a yield of 1.5-2.5 ug per extract.

[ATϟGC]

I'll echo ECO and melop in saying that I've tried SPRI beads (Ampure and Seramag speedbeads) to clean up WGA products as well as genomic DNA from home-brew library prep and genomic DNA extraction protocols. As ECO mentioned, they do tend to clump and stick together with higher molecular weight fragments. It might help slightly if you avoid drying the bead pellet till it cracks. Longer elution times (I often incubate overnight at 4 Celsius for genomics extracts) and a small amount of Tween-20 seems to help elute more of the bound DNA.

[melop]

It works for genomic DNA. I now use SPRI beads for extracting genomic DNA either directly from proK-SDS digested tissue or from the supernatant cleaned by phenol-chloroform.

[clintp]

Very cool--do you actully use them for WGA products?

[ECO]

HMW products will bind just fine, in fact you'll see the beads clump dramatically with WGA products. You will have problems eluting them so incubate for a while, pipet a bunch and/or vortex.

[robertami]

I've never seen fragments >700 bp with Ampure beads, I guess you could try lowering the Beads: DNA ratio (titrate maybe?) and run the elute on a gel and check the max size you manage to get

[clintp]

Thanks for the reply, but that's not exactly what I'm asking. Since the beads are typically used for PCR cleanup and size-selection of small fragments, I wondered if larger molecules of DNA, like 15-100kb, are efficiently bound to the beads and are efficiently eluted.