Gel size selection for next generation sequencing (NGS) fragment library preparation is the standard process used in laboratories to achieve size selection. During this tedious process, time needs to be spent purifying required fragment sizes from the gel, which can only result in a low throughput. To this end, scientists from Beckman Coulter have launched the SPRIselect® reagent kit to provide a high throughput, streamlined and automation-friendly method for size selection.
SPRIselect is a paramagnetic bead technology based on solid phase reversible immobilization (SPRI) and can be used in small to medium throughput workflows and adjusted for various sequencing applications. The beads selectively bind fragments based on the SPRIselect reagent:sample ratio. This ratio can be adjusted to selectively keep or discard fragment sizes that lie outside of the desired range. The adjustable workflow for the kit enables users to carry out, right, left, or double size selection. Here, the different workflows are described to show how SPRIselect beads work as the reagent:sample ratio is altered.
For left side selection, the appropriate ratio volume of SPRIselect reagent is added to the sample. The beads bind to larger fragments to the right of the target range and are eluted. Smaller fragments left of the target range are retained in the supernatant and can be discarded or retained.
For right side selection, the appropriate ratio of beads are added, which binds larger fragments right of the target range that are to be discarded, while smaller fragments left of the range are removed in the supernatant to a fresh tube. Using a re-bind step, the buffer is exchanged by re-adding beads to the supernatant to bind all fragments.
For double size selection, the same workflow for right side selection can be used, but the re-bind step is used to perform left side selection. The volume of beads required to perform a left side selection re-bind is calculated by subtracting the right side ratio from the left side ratio and multiplying by the volume of supernatant that was transferred.
Gel-free size selection provides significant time savings by eliminating the need to perform difficult purification steps. To save even more time, users can automate size selection by defining different size selection ratios for each well using a .CSV file. A simple user interface enables the operator to customize selection type, sample input volume and binding and elution parameters.
For further information on the SPRIselect reagent kit, please click here.
About Beckman Coulter
Beckman Coulter develops, manufactures and markets products that simplify, automate and innovate complex biomedical tests. More than a quarter of a million Beckman Coulter instruments operate in laboratories around the world, supplying critical information for improving patient health and reducing the cost of care.
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