ROCHE’S KAPA EVOPLUS BOOST KIT (RUO)

Highly ReproducibleIllumina Libraries

Volta’s App on Callisto for Roche’s KAPA EvoPlus Boost Kit (RUO)* enables an automated and streamlined workflow for converting DNA inputs of varying quantities into Illumina™-compatible, sequencing-ready libraries. The workflow integrates key library preparation steps—including enzymatic fragmentation & A-tailing, adapter ligation, and bead-based size selection— within a single automated process.

key benefits:

Consistent and tunable fragmentation
Reproducible insert sizes across DNA inputs and GC content enableconsistent, high-quality libraries.
Improved sequencing efficiencydelivers highly uniform coverage Reduced sequencing artifactsand low duplication rates (<10%)maximize usable sequencing reads.
Streamlined, walk-awayautomation on Callisto Combined fragmentation andA-tailing ReadyMix chemistrysimplifies setup and enables robust,automated library preparation withminimal hands-on time.

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Built on the proven performance of KAPA library preparation chemistry, the workflow delivers reduced sequencing artifacts and robustness to common inhibitors. Combined fragmentation and A-tailing in ReadyMix formulations simplify workflow setup and enable robust library preparation. The App supports a range of sequencing applications, including whole genome sequencing (WGS) and whole-exome sequencing (WES) and/or targeted sequencing.

App Specifications

Input: 100-500 ng DNA

Output: Sequencing ready libraries

Hands-on time: Less than 30 minutes

Walk-away time: 3 hours

Throughput: Flexible, 4 - 24 rxns in a single run

Chemistry: Roche KAPA EvoPlus Boost(PCR-free) Kit (RUO)

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Highly reproducible output library size across DNA inputs

The library prep App consistently generates libraries with a mode fragment size of ~540 bp across DNAinputs ranging from 100 ng to 500 ng, producing libraries suitable for whole-genome sequencing.

Robust library yield across a wide DNA input range

Library concentrations exceeding 2 nM were obtained across DNA inputs from 100 ng to 500 ng,demonstrating robust performance across a wide range of sample inputs. DNA concentration wasquantified using Qubit and TapeStation.

Consistent performance across DNA sources

The App supports diverse DNA sources, including buffy coat, human reference DNA (HG001),and DNA extracted from whole blood (WB), producing consistently high library concentrations.DNA concentration was quantified using Qubit and TapeStation.

High accuracy variant calling

Automated library preparation on Callisto delivers variant calling accuracy comparable to or betterthan manual workflows. Libraries prepared on Callisto (n=7 technical replicates) demonstrated higherSNP recall, precision, and F1 scores compared to manually prepared libraries (n=5), with comparableperformance for indel detection. Libraries were generated using HG001 DNA, sequenced on theIllumina NovaSeq X, and benchmarked against the NIST reference set (HG001_GRCh38_1_22_v4.2.1_benchmark.bed) using hap.py.