Biomics

Introduction. High-throughput DNA sequencing is currently a standard part of cancer treatment, both for treatment diagnosis and early-stage screening. NGS dominates among the sequencing methods, and it has a number of significant drawbacks. The evolving nanopore sequencing technology overcomes all of the above limitations of NGS. The accuracy of such sequencing is closer to NGS, and the technology itself is more affordable in terms of budget. Nanopore sequencing has already found wide application in various applications and there is a significant prospect of its application in polymorphism analysis to identify genetic variations for diagnostic and prognostic tasks.

The purpose of the study: based on the developed amplicon panels for the detection of single nucleotide polymorphisms on the example of DNA lymphoma samples, to conduct a study on optimizing the amplification process, nanopore sequencing and data analysis using a domestic reagent database and a domestic nanopore sequencer "Nanoporus Pro".

Materials and methods. The DNA isolated and purified using spin columns from paraffin sections was amplified using selected primers (47 pairs) in a 4-set multiplex. The resulting DNA libraries were sequenced using the nanopore sequencer Nanoporus Pro.

Results and conclusion. Multiplex PCR conditions based on several variants of domestic DNA polymerases were investigated and optimized. During the analysis of the obtained sequencing data, 45 out of 47 reference targets were identified and highly accurate data on the presence of single nucleotide polymorphisms at the studied points of variation were obtained. Thus, the high prospects of using nanopore sequencing for multiplex analysis of clinically important mutations in cancer genome regions using domestic reagents and equipment have been shown.

nanopore sequencing, NGS, single nucleotide polymorphisms, lymphomas

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