Temperature sensitivity and reagent handling FAQ for TruSight Oncology 500 (TSO500) ctDNA

Is there any range of temperature sensitivity for the room temperature incubations during the enrichment steps? All testing was performed in a controlled lab environment with temperatures typically ranging from 20-25°C. That said, some variation is likely acceptable within reason for room temperature. Illumina recommends avoiding bench tops that are in direct sunlight or performing testing in a space that is not temperature controlled (eg, lab spaces without heating or air conditioning).

Can reservoirs and multichannel pipettors be used for library prep? Will the provided reagent volume be sufficient if reservoirs are used? Are there any particular handling techniques that are different from the TSO500 Tissue protocol? Yes, sterile reservoirs and multichannel pipettors can be used for the TSO500 ctDNA workflow. The kit reagents are sufficient for 48 libraries, though this does not include overage specific for reservoir use, as volumes needed can vary. For small volumes (such as enzyme mixes, etc.) volumes can be pipetted into strip tubes for multichannel dispensing. Reagent troughs can be used for bead steps and HP3+EE2 master mixes, but be cautious to not overuse reagents.

What is the time sensitivity of HP3+EE2 master mix? Follow the handling instructions in the reference guide. The elution master mix (HP3+EE2) is not a sensitive reagent, and can be stable at room temperature for up to 30 minutes.

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