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.
ФГБНУ «Центральный НИИ туберкулеза», Москва, РоссияЦель исследования: установить виды нетуберкулезных микобактерий (НТМ), выделяемых из дыхательных путей больных в различных регионах РФ, используя для идентификации НТМ ДНК-стрипы�
MoniToring oF sPecies diVersiTY oF non-Tuberculosis MYcobacTeria in THe soMe russian regions using dna-sTriPs oF genoTYPe MYcobacTeriuM cM/as (Hain liFescience, gerManY)
T. G. SMIRNOVА, S. N. АNDREEVSKАyA, E. E. LАRIONOVА, I. yU. АNDRIEVSKАyA, V. V. USTINOVА, L. N. CHERNOUSOVА central Tuberculosis research institute, Moscow, russiaGoal of study: to define species of non-tuberculosis mycobacteria, isolated from the respiratory tract of the patients from various Russian regions using DNA strips for identification of non-tuberculous mycobacteria� Materials and methods. 1204 cultures of non-tuberculosis mycobacteria isolated on solid and liquid media from 727 patients in whom tuberculosis/mycobacteriosis was suspected were examined from 2011 to February, 2017� Species of non-tuberculosis mycobacteria were identified using DNA-strips of Hain Lifescience (Germany) GenoType ® Mycobacterium CM/AS in compliance with manufacturer's instructions� results. 17 species of non-tuberculosis mycobacteria were identified (11 species belonged to the group of those slow-growing and 6 species belonged to fast growing non-tuberculosis mycobacteria)� Slow-growing mycobacteria prevailed (564/727, 77�58%) and M. avium was the most prevalent (210/727; 28�89%)� The following mycobacteria were found (in descending order): M.
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