Fully-Automated Cross-Platform DNA Sample Preparation for Next-Generation Sequencing

The Next-Generation sequencing platforms by Life Tech (SOLiD), Illumina (Hiseq 2000), Roche (454 Titanium) and Ion Torrent (PGM), can generate 100gbs of data. The bottleneck is no longer in the sequencing, but in Library preparation and data analysis. The PSS 8G Compact Automated Library Preparation System and Reagent offering works to eliminate the library preparation bottleneck.

Jeff Ta1, Wilson Liu1, James Pass2, Morten Lukacs3, Anders Holmberg3,
Akram Tariq1, Carl Hilliker2, Nader Pourmand1

Exploiting the well recognized phenomenon of
DNA breakage caused by a transition metal
in combination with a reducing agent, we optimized
conditions to obtain DNA fragment sizes anywhere
between 50bp - 1000bp for DNA / RNA fragment
libraries and 2kb-10kb for mate-pair libraries.
Minimal base damage is caused by this fragmentation
method as comared with conventional DNA
shearing technology.

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Procedure Overview

Reduce risk of human error
Prevent cross contamination
Increase reproducibility

Provide ease of handling
Scalability
Minimize cost

1 University of California Santa Cruz CA
2 Precision System Science USA CA
3 NorDiag ASA Norway

Effect of different incubation times
using equimolar concentration of a transition metal in combination with a
reducing agent to produce DNA
fragmentation.

Metal Based Fragmentation System

METAL INDUCED OXIDATIVE
DNA BREAKAGE

Introduction

Advantages to Automation