AMPure XP Beads for DNA Cleanup
Applications: PCR cleanup, DNA sample concentration, NGS library prep
The gold standard in bead-based, next-generation sequencing (NGS) cleanup—in fact, it's suggested in over 200 library preparation kits from trusted sequencing companies.
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Overview
The AMPure XP bead-based reagent is intended for performing DNA cleanup steps in different genomic applications like sequencing, qPCR/ddPCR/PCR, microarrays, and other enzymatic reactions. It is powered by proven SPRI technology, which uses paramagnetic beads to selectively bind nucleic acids by type and size.
Works with DNA
High recovery of amplicons, greater than 100 bp
Efficient removal of unincorporated dNTPs, primers, primer dimers, salts and other contaminants
Manual or automated to fit any throughput
Maximizing recovery, consistency, and speed to facilitate the entire NGS workflow, the AMPure XP reagent meets the stringent needs of today’s genomic applications and minimizes the risk of losing important genetic information. That's why it's suggested in over 200 NGS library preparation kits often as the sole cleanup option, including those from the industry's most trusted sequencing companies like Illumina, Oxford Nanopore Technologies, PacBio and Thermo Scientific.
AMPure XP DNA Cleanup Workflow and Protocol
- Bind DNA to AMPure XP beads
- Separate beads from contaminants
- Wash beads with 70% ethanol to remove contaminants
- Elute DNA from AMPure XP beads
- Transfer to new tube or plate
Specifications
| Nucleic Acid Input | PCR products, fragmented DNA |
| Output | Purified and/or concentrated DNA |
| Recovery | AMPure XP beads were developed for the purification of PCR products 100bp<. Normal recovery ranges from 60-90%, and is typically between 70-90%. |
| Bead Ratio | 1.8X for cleanup |
| Format | Liquid |
| Volumes | 5 mL, 60 mL, or 450 mL |
| Applications | PCR cleanup, DNA sample concentration, NGS library prep |
| Processing mode | Automated or manual |
| Technology | SPRI paramagnetic bead-based technology |
| Storage | 4 °C |
How to Automate AMPure XP DNA Cleanup
When compared to manual operations, the AMPure XP beads automated on Biomek liquid handlers provides:
- Reduced hands-on time and increased throughput
- Option to run the method end-to-end with only setup and tear-down touch points
- Reduction in pipetting errors
- Standardized workflow for improved results
- Quick implementation with demonstrated methods
To learn more about automation of the AMPure XP beads on the Biomek i5 Nucleic Acid Cleanup Solution, read our method overview.
To discuss the feasibility to automate the AMPure XP DNA cleanup in your lab with one of our experts complete this form.
Frequently Asked Questions
Although size selection protocols developed by other organizations do exist, we cannot guarantee performance or support this application. We suggest using the SPRIselect reagent, as it is developed specifically for size selection purposes. Visit this page to learn more.
The main difference between the SPRIselect and AMPure XP bead-based reagents is the intended use. The SPRIselect reagents are designed and validated for accurate and consistent DNA size selection from lot to lot, while the AMPure XP reagents are primarily validated for PCR purification and NGS cleanup. Learn more on this page.
The reagent will bind DNA and RNA; however, the RNAClean XP reagent is manufactured under RNase-free conditions and is QC tested to be RNase free whereas the AMPure XP reagent is not.
The AMPure XP reagent is manufactured and tested for the storage temperature indicated on the bottle and we can guarantee performance only at that temperature.
No. Any beads carried over to the final destination plate are inert in downstream enzymatic reactions.
Yes, but the recovery will decrease with a decrease in elution volume. In addition, if the beads cannot reach the magnet during the separation it may be necessary to leave a portion of the eluate behind if bead carryover must be avoided. Be sure to resuspend the beads fully in the elution buffer.
The binding capacity of the beads is so high that they cannot be exceeded with any sample type. It is possible to bind at least 7 μg of nucleic acid to 1 μL AMPure XP reagent, however the sample will become so viscous that it is difficult to pipette the sample.
The recovery depends on the fragment size, sample volume, sample concentration, elution volume and some other factors. Under ideal conditions recovery can be up to 90%, but under sub-optimal it can be as low as 60%.
Our beads rely upon a multifactorial equilibrium chemistry for binding, washing, and elution. Binding buffer components facilitate nucleic acid immobilization at functional-group-rich binding sites on the beads and the wash steps preserve this delicate equilibrium while solubilizing contaminants to allow for their removal. Elution disrupts this balance and re-solubilizes nucleic acids to allow for their separation from the beads.
Learn More →
A dry step can be incorporated after the removal of the last ethanol wash and before the addition of the elution buffer. For purification of very large fragments, such as gDNA, a dry step is not recommended because recovery would be decreased. Fragments up to 40 kb have been known to tolerate a dry step.
Find Beckman Coulter Life Sciences AMPure XP technical documents here.
Didn't find an answer to your question? Talk to our expert.
Citations
AMPure XP beads have been cited in over 65,000 publications on Google Scholar and referenced in articles in Science, Nature, and PNAS. Here are some featured articles:
Frequently Used With
SPRIPlate, 96-well, Super Magnet
Specifically designed for SPRI reagent kits by Beckman Coulter. Has not been validated for use with the CleanSEQ or CosMCPrep reagent systems.
SPRIStand, Tubes
Designed for lower throughput users. The 6-position stand is compatible with 1.5 mL, 1.7 mL and 2.0 mL microcentrifuge tubes.
SPRIselect Bead-Based Reagent
SPRIselect beads utilize SPRI technology for DNA size selection and cleanup and to deliver superior quality DNA that requires no centrifugation or filtration.
DNA Extraction
Beckman Coulter's DNA extraction systems and accessories are based on SPRI paramagnetic bead-based technology in an automation-compatible format. Superior DNA isolation with no salt carry over. Better isolate DNA with magnetic SPRI bead technology.
Technical Documents
Products and demonstrated applications are not intended or validated for use in diagnostic procedures.
