Added values

MASTR Solutions

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Increasing output and reducing workload with Next-Generation Sequencing technologies

Multiplicom N.V. [ a ] Galileïlaan 18, B-2845 Niel, Belgium [ w ] www.multiplicom.com [ e ] customerservice@multiplicom.com [ t ] +32 3 289 14 00 [ f ] +32 3 400 23 16

Enhanced target amplification

MASTR assays offer an innovative combination of premixed PCR primers in a ready to use kit,

enabling enhanced target amplification of DNA.

Multiplicom’s MASTR (Multiplex Ampli-fication of Specific Targets for Resequencing) assays enable multiplex PCR amplification of all required coding sequences of the genes of interest in a limited number of PCR reactions. Further downstream pooling of DNA amplicons and barcoding individual samples of the MASTR assays with contemporary Next-Generation Sequencing (NGS) technologies, allows simple, high throughput and cost-effective sequencing for both research and diagnostic purposes.

MASTR assays serve as front-end amplification for sequence analysis on all NGS systems. The technology is based on “target amplification“ method rather than target enrichment. Target gene selection for all MASTR assays, based on

input from key opinion leaders, as well as Multiplicom’s proprietary primer design algorithm and know-how guarantees in a highly reproducible simultaneous PCR amplification of several 100 amplicons within a single, standard PCR reacction. As a result, MASTR assays enable more accurate runs, at significantly higher throughputs and substantially reduced workload and sample processing costs, using NGS technologies.

The MASTR assays employ a simple 2-step PCR protocol enabling specific amplification of the regions of interest (step 1) followed by the incorporation of molecular barcodes (Multiplex Identifiers - MIDs) in each amplified product (step 2) to unambiguously link each read to the sample it originated from.

View of the two-step amplification process involved in the MASTR assay technology.

Step 2

Step 1

Based on a 2-step simple PCR protocol

MASTR assays offer an innovative combination of premixed PCR primers in a ready to use kit,

enabling enhanced target amplification of DNA.

A MASTR assay is a simple process, consisting of two steps:

First StepMultiplex PCR amplification of target of interest (e.g. BRCA, CFTR, EGFR MASTR assays).

Second Step: For long reads sequencers: Flexible barcoding incorporation in each amplified product to unambiguously link each read to the original sample (e.g. drMID kit for long reads up to 500 bp).

For short reads sequencers: Second PCR amplification and cleanup to reduce the amount of excess primers (e.g. Short Read Amplification kit for short reads from 50-200 bp).

This is followed by a fixed scheme to mix multiplexes, enable PCR clean-up and for equimolar pooling of barcoded libraries.

Optional QC step: After the second step you can perform fragment analysis via fluorescence labeling. Data can be analyzed with the MAQ-S analysis software, which you can download from our website (www.agilent.com).

Important note: For “short read” sequencing technologies, you have the option to perform the QC step mentioned above before proceeding with the short fragment generation steps by purification, concatenation, enzymatic shearing and / or adaptor ligation kits.

A simple, easy-to-use workflow

Multiplex PCR Universal PCR SEQUENCING

DATA ANALYSIS

Pooling & Clean up

NGS

Sample Preparation

Quality Control

Ready to run amplification • Ready-to-use kit comprising full coding regions• Greater target specificity and superior spread factor• Solution for homopolymer stretches and copy number variations

High throughput • Flexible assay design• Leveraging NGS capacity: uniform representation of all amplicons at a narrow range• Identical protocol among all MASTR assays

Convenient & compatible workflow • Standard PCR protocol and equipment, with minimum hands on time• Compatible with Illumina NGS systems and Life Technologies Ion NGS Systems• Data analysis support

Added values

v.1707 - EUFor Research Use Only. Not for use in diagnostic procedures.

Multiplicom N.V. [ a ] Galileïlaan 18, B-2845 Niel, Belgium [ w ] www.multiplicom.com [ e ] customerservice@multiplicom.com [ t ] +32 3 289 14 00 [ f ] +32 3 400 23 16