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Comparison of whole genome amplification techniques for human single cell exome sequencing.

ABSTRACT:

Background

Whole genome amplification (WGA) is currently a prerequisite for single cell whole genome or exome sequencing. Depending on the method used the rate of artifact formation, allelic dropout and sequence coverage over the genome may differ significantly.

Results

The largest difference between the evaluated protocols was observed when analyzing the target coverage and read depth distribution. These differences also had impact on the downstream variant calling. Conclusively, the products from the AMPLI1 and MALBAC kits were shown to be most similar to the bulk samples and are therefore recommended for WGA of single cells.

Discussion

In this study four commercial kits for WGA (AMPLI1, MALBAC, Repli-G and PicoPlex) were used to amplify human single cells. The WGA products were exome sequenced together with non-amplified bulk samples from the same source. The resulting data was evaluated in terms of genomic coverage, allelic dropout and SNP calling.

SUBMITTER: Borgstrom E

PROVIDER: S-EPMC5313163 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Comparison of whole genome amplification techniques for human single cell exome sequencing.

Borgström Erik E Paterlini Marta M Mold Jeff E JE Frisen Jonas J Lundeberg Joakim J

PloS one 20170216 2

<h4>Background</h4>Whole genome amplification (WGA) is currently a prerequisite for single cell whole genome or exome sequencing. Depending on the method used the rate of artifact formation, allelic dropout and sequence coverage over the genome may differ significantly.<h4>Results</h4>The largest difference between the evaluated protocols was observed when analyzing the target coverage and read depth distribution. These differences also had impact on the downstream variant calling. Conclusively, ...[more]

PMID: 28207771

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