Details

Autor(en) / Beteiligte

• Picher, Ángel J
• Budeus, Bettina
• Wafzig, Oliver
• Krüger, Carola
• García-Gómez, Sara
• Martínez-Jiménez, María I
• Díaz-Talavera, Alberto
• Weber, Daniela
• Blanco, Luis
• Schneider, Armin

Titel

TruePrime is a novel method for whole-genome amplification from single cells based on TthPrimPol

Ist Teil von

• Nature communications, 2016-11, Vol.7 (1), p.13296-13296, Article 13296

Ort / Verlag

England: Nature Publishing Group

Erscheinungsjahr

2016

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Sequencing of a single-cell genome requires DNA amplification, a process prone to introducing bias and errors into the amplified genome. Here we introduce a novel multiple displacement amplification (MDA) method based on the unique DNA primase features of Thermus thermophilus (Tth) PrimPol. TthPrimPol displays a potent primase activity preferring dNTPs as substrates unlike conventional primases. A combination of TthPrimPol's unique ability to synthesize DNA primers with the highly processive Phi29 DNA polymerase (Φ29DNApol) enables near-complete whole genome amplification from single cells. This novel method demonstrates superior breadth and evenness of genome coverage, high reproducibility, excellent single-nucleotide variant (SNV) detection rates with low allelic dropout (ADO) and low chimera formation as exemplified by sequencing HEK293 cells. Moreover, copy number variant (CNV) calling yields superior results compared with random primer-based MDA methods. The advantages of this method, which we named TruePrime, promise to facilitate and improve single-cell genomic analysis.