Question: What are the best practices for using SPRIselect beads in 10x assays?
Answer: Due to the sensitivity of the size selection to the SPRIselect: DNA ratio, minor differences in SPRIselect bead volume can shift the peak of the size distribution trace either to smaller or larger fragment sizes. Therefore, a consistent technique is essential, and the following guidelines are suggested to maintain uniform fragment size distributions.
- Ensure SPRI beads are not expired
- Use well-calibrated pipettes
- Use appropriate pipette for SPRI beads handling to increase pipetting accuracy (e.g., P200 for volumes of 30 µl - 200 µl)
- Ensure that SPRI beads are fully thawed and equilibrated to room temperature before use
- Vortex SPRI beads thoroughly before each transfer to sample
- Avoid transferring excess SPRI beads that may have been retained on the outside of the pipette tip or as a drop hanging from the tip
- While doing ethanol washes and while eluting samples off the beads, visually inspect the solutions in the tips to ensure that the beads are not accidentally aspirated
- Mix SPRI beads thoroughly into solutions to create a uniform mixture
- Always use fresh preparations of 80% Ethanol
- Ensure not to overdry the beads pellet