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, 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 prior to use
- Vortex SPRI beads thoroughly prior to each transfer to sample
- Avoid transferring of excess SPRI beads that may have retained on the outside of the pipette tip or as a drop hanging from the tip
- Visually inspect the levels of solutions in tips during dispensing to ensure the beads are not accidentally aspirate
- 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