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ISSN 2073-8137
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Development of the method for analysis of bacillus anthracis single nucleotide polymorphisms using the mini sequensing and mass spectrometry

[Experimental medicine]
Evgeniy Eremenko; Vladimir Voropayev; Elena Kotenyova; Alla Ryazanova; Lyudmila Aksyonova; Olga Tsygankova; Nina Buravtseva; Alexandr Kulichenko;

Objective: development of the method for the B. anthracis SNP-genotyping using the mini sequencing and MALDI TOF MS. Material and methods: DNA of B. anthracis strains, oligonucleotides mass specters using regents for PCR, mass spectrometry, DNA amplifier, MALDI TOF mass spectrometer, methods for PCR amplification, DNA sequencing and
MALDI TOF mass spectrometry, computer hardware, BLASTn on line resource and GenBank data base were used. Results. Primers for the 13 B. anthracis canonical SNPs mini sequencing and nucleoside triphosphates composition were designed, parameters for mini sequencing reaction were developed, reproducibility and correctness of obtained results by methods of real time allele-specific PCR amplification and direct sequencing were checked. Conclusion: method for the SNP-genotyping using the mini sequencing and MALDI TOF MS has been developed.

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References:
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Keywords: B. anthracis, genotyping, mini sequencing, MALDI TOF mass spectrometry


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