PT  - JOURNAL ARTICLE
AU  - Konstantin A. Blagodatskikh
AU  - Vladimir M. Kramarov
AU  - Ekaterina V. Barsova
AU  - Alexey V. Garkovenko
AU  - Dmitriy S. Shcherbo
AU  - Andrew A. Shelenkov
AU  - Vera V. Ustinova
AU  - Maria R. Tokarenko
AU  - Simon C. Baker
AU  - Tatiana V. Kramarova
AU  - Konstantin B. Ignatov
TI  - Improved DOP-PCR (iDOP-PCR): a robust and simple WGA method for efficient amplification of low copy number genomic DNA
AID  - 10.1101/128736
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 128736
4099  - http://biorxiv.org/content/early/2017/04/19/128736.short
4100  - http://biorxiv.org/content/early/2017/04/19/128736.full
AB  - Whole-genome amplification (WGA) techniques are used for non-specific amplification of low-copy number DNA, and especially for single-cell genome and transcriptome amplification. There are a number of WGA methods that have been developed over the years. One example is degenerate oligonucleotide-primed PCR (DOP-PCR), which is a very simple, fast and inexpensive WGA technique. Although DOP-PCR has been regarded as one of the pioneering methods for WGA, it only provides low genome coverage and a high allele dropout rate when compared to more modern techniques. Here we describe an improved DOP-PCR (iDOP-PCR). We have modified the classic DOP-PCR by using a new thermostable DNA polymerase (SD polymerase) with a strong strand-displacement activity and by adjustments in primers design. We compared iDOP-PCR, classic DOP-PCR and the well-established PicoPlex technique for whole genome amplification of both high- and low-copy number human genomic DNA. The amplified DNA libraries were evaluated by analysis of short tandem repeat genotypes and NGS data. In summary, iDOP-PCR provided a better quality of the amplified DNA libraries compared to the other WGA methods tested, especially when low amounts of genomic DNA were used as an input material.