SPRIselect Bead-Based Reagent

Applications: DNA Size Selection, PCR Purification, NGS Cleanup

Suggested in over 40 NGS library preparation kits, the SPRIselect reagent provides flexibility and control over the DNA size selection process with minimal lot-to-lot variance. Plus, it's stable at room temperature.

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Overview

The SPRIselect bead-based reagent is designed to provide scientists with more control over the size selection step during NGS library preparation, which is critical for generating high-quality sequencing data. It is powered by proven SPRI technology, which uses paramagnetic beads to selectively bind nucleic acids by type and size.

Works with fragmented DNA

Tunable from 150 to 800 bp to offer easy adjustments for specific applications and sequencers

Eliminates the needs for gels, chips or special instrumentation

High level of predictability and consistency between different runs and reagent lots

The simplified workflow and automation capability of SPRIselect reagents make the process of size selection more streamlined and less prone to errors, resulting in faster and more reliable sequencing results.

Specifications

Nucleic Acid Input Fragmented DNA, PCR products
Output Size Selected DNA
Range of Size Selection 150–800 bp
Bead Ratio Size Selection: Depends on selection point(s), Cleanup: 1.8X.
Format Liquid
Volumes 5 mL, 60 mL, or 450 mL
Applications DNA Size Selection, PCR Purification, NGS Cleanup
Processing mode Automated or manual
Technology SPRI paramagnetic bead-based technology
Storage Room Temperature

Data and Performance

Two different size selections were performed on sheared gDNA from E. coli using SPRISelect reagent. Triplicates were performed per size selection. Each of the triplicates show very consistent size selection capabilities. (Left) Bind to sample ratio of 0.5X for left side size selection. (Right) Bind to sample ratio of 0.5X for right side size selection. DNA size was measured using an Agilent 2100 Bioanalyzer high sensitivity chip. The peaks at 43 and 115 are internal markers.

SPRIselect Beads—Left Side DNA Size Selection Performance

SPRIselect Beads—Right Side DNA Size Selection Performance

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Workflows and Protocol

SPRIselect Left Side Size Selection Workflow

SPRIselect Beads—Left Side Size Selection Workflow
  1. Bind DNA to magnetic beads
  2. Separate beads from contaminants
  3. Wash magnetic beads with 85% ethanol to remove contaminants
  4. Elute DNA from magnetic beads
  5. Transfer to new plate

SPRIselect Right Side Size Selection Workflow

  1. Bind DNA to magnetic beads
  2. Transfer supernatant to a new plate
  3. Bind DNA to magnetic beads
  4. Separate beads from contaminants
  5. Wash magnetic beads with 85% ethanol to remove contaminants
  6. Elute DNA from magnetic beads
  7. Transfer to new plate

SPRIselect bead-based reagent can be performed either manually or automated on a liquid handling system depending on batch size or overall throughput.

In the table you can see estimated hands-on time and total time, in minutes, required to perform 8, 24, 48 and 96 DNA size selections on the Biomek i7 Hybrid Workstation using SPRIselect reagent.

To learn more about automating DNA size selection on the Biomek i-Series Workstations, read our application note.

Read the Application Note

Manual Workflow Automated Workflow
Batch Size 8 Hands-on Time 10 10
Total Time 20 40
24 Hands-on Time 15 10
Total Time 25 42
48 Hands-on time NR 15
Total Time NR 51
96 Hands-on time NR 15
Total Time NR 60

NR = Not Recommended

Resources

Brochure

 

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Data Sheet

 

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Customer Spotlight

 

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Customer Spotlight

 

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Application Note

 

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Brochure

 

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Frequently Asked Questions

The main difference between the 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. Read more about the differences here.

We cannot guarantee post-thaw suitability or stability for every application.

No. Any beads carried over to the final destination plate are inert in downstream enzymatic reactions.

We recommend 40 μL. The beads should be submerged when they are in the elution buffer, otherwise when you transfer the eluate to another plate, you may get excessive bead carryover or low yield. In most 96 well PCR plates, it takes 40 μL to cover the beads. Certain combinations of third-party labware and magnetic separation plates may allow for lower elution volumes.

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.

Recovery will vary based on the range of fragment sizes selected and is heavily dependent on the input sample. In general, as specificity increases (and the fragment size range selected narrows) yield will also decrease.

Find Beckman Coulter Life Sciences SPRIselect technical documents here.

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Citations

SPRIselect reagents have been cited in over 3,000 publications on Google Scholar and referenced in articles in Science, Nature, and PNAS. Here are three featured articles:

Size Selection of Libraries
Greenwald WW et al. Subtle changes in chromatin loop contact propensity are associated with differential gene regulation and expression. Nat Commun. 2019 Mar 5;10(1):1054. doi: 10.1038/s41467-019-08940-5.
Used on cDNA Libraries
Kubo M et al. Single-cell transcriptome analysis of Physcomitrella leaf cells during reprogramming using microcapillary manipulation. Nucleic Acids Res. 2019 May 21;47(9):4539-4553. doi: 10.1093/nar/gkz181.
Used on ChIP-seq Libraries
Behera V et al. Interrogating Histone Acetylation and BRD4 as Mitotic Bookmarks of Transcription. Cell Rep . 2019 Apr 9;27(2):400-415.e5. doi: 10.1016/j.celrep.2019.03.057.

Documents techniques

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Products and demonstrated applications are not intended or validated for use in diagnostic procedures.