SPRISelect Size Selection Performance and Data

SPRISelect Size Selection Solution Performance & Data

Next Generation Sequencing (NGS) is the most pervasive technology in genomics research today. The long list of front-end applications has varied library preparation requirements that demand flexible solutions. The size of DNA has a direct impact on generating desired sequencing results. The SPRISelect Reagent Kit is designed to give scientists more flexibility and control over the size selection process. With proven, industry leading Solid Phase Reversible Immobilization (SPRI) technology, this reagent provides robust, reproducible and customizable size selection.

  • Tunable from 150 to 800 base pairs allows for easy adjustments for specific applications and sequencers
  • Efficient mag bead technology eliminates the need for gels, chips or special instrumentation
  • Scalable from manual to automated for high throughput processing in 96-well plates

 

Dual Size Selection Capability

Left size selection removes DNA that is smaller than the targeted size, and right size selection removed DNA that is larger than the target size. By utilizing both left and right size selection you can target a smaller range of DNA sizes.

SPRISelect-left size selctionSPRISelect-right size selection

 

SPRISelect-overlay of left and right size selectionsSPRISelect-double size selection

 

SPRISelect-overlay of left and right size selections

 

SPRISelect was used for dual size selection on sheared gDNA from E. coli. [Figure A] Left size selection with a ratio for 400bp. [Figure B] Right size selection with a ratio for 200bp. [Figure C] An overlay of the left size selection (red) and right size selection (blue). [Figure D] Double size selection excludes fragments above and below target cutoffs and captures a targeted regions between 200bp and 400bp. [Figure E] Sheared gDNA from E. coli. For all DNA size was measured using an Agilent 2100 Bioanalyzer high sensitivity chip (Agilent). The peaks at 35 and 10300 are internal markers.

 

Reproducible and Tunable

SPRISelect is manufactured to tight specifications ensuring run to run and lot to lot reproducibility.

Two different size selections were performed on sheared gDNA from E. coli using SPRISelect. 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 (Agilent). The peaks at 43 and 115 are internal markers.

SPRISelect-Size selection ration of 0.5xSPRISelect-E coli size selection-ratio 0.5x

 

(Left) Sizes election ratio of 0.5x. (Right) Size selection ratio of 0.5x.

 

Less Hands-On Time Than Other Suppliers

SPRISelect and two other commercially available kits were used for size selection on sheared gDNA from E. coli. (Left) The graph represents the time for single size selection or clean up. The SPRISelect workflow for a single size selection is 4.2 and 2 times faster than supplier A and supplier B, respectively. (Right) The graph represents the time for dual size selection. The SPRISelect workflow for dual size selection is 3.8 and 2 times faster than supplier A and supplier B, respectively. SPRISelect has the lowest processing time out of all three suppliers by at least 50%. The times were based on performing size selection for 8 samples manually. Genomics SPRISelect Cleanup Time Genomics SPRISelect Size Selection Time

 

SPRIselect Left Workflow

Selecting for DNA larger than a target size.

Genomics SPRIselect 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 a storage plate

 

SPRIselect Right Workflow

Selecting for DNA smaller than a target size or selecting for DNA within a target region.

Genomics 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 a storage plate

 

Simple to Run Manually or Automated

Estimated hands-on time and total time in minutes, required to perform dual size selection on 8, 24, 48 and 96 DNA samples using SPRISelect. The different methods can be performed either manually or automated on a Biomek workstation. The table represents automation times performed on a Biomek i7 hybrid. Difference in time between manual and automation is indicated. NR=Not Recommended.

SPRISelect Manual Automated
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

 

Products for Size Selection

PART # NAME PREPS
B23317 SPRISelect Reagent 5 mL
B23318 SPRISelect Reagent 60 mL
B23319 SPRISelect Reagent 450 mL

 

 

Not intended or validated for use in the diagnosis of disease or other conditions.

Copyright 2019 Beckman Coulter, the stylized logo, and the Beckman Coulter product and service marks mentioned herein are trademarks or registered trademarks of Beckman Coulter, Inc. in the United States and other countries. All other trademarks are the property of their respective owners.



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