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Hematology - Oncology

A Modified Method for Whole Exome Resequencing from Minimal Amounts of Starting DNA
Published: Monday, March 05, 2012
Author: Iwanka Kozarewa et al.

by Iwanka Kozarewa, Juan Manuel Rosa-Rosa, Christopher P. Wardell, Brian A. Walker, Kerry Fenwick, Ioannis Assiotis, Costas Mitsopoulos, Marketa Zvelebil, Gareth J. Morgan, Alan Ashworth, Christopher J.

Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes.