scbmtБИОМЕДИЦИНАJournal Biomed2074-59822713-0428Scientific center of biomedical technologies of Federal Medical and Biological Agency10.33647/2713-0428-17-3E-48-52scbmt-1311Research ArticleМЕТОДЫ И ТЕХНОЛОГИИ БИОМЕДИЦИНСКИХ ИССЛЕДОВАНИЙMETHODS AND TECHNOLOGIES OF BIOMEDICAL RESEARCHГенотипирование линий мышей по гену TPH2 в рутинной практике: выбор методики выделения ДНК для проведения ПЦРGenotyping of mice lines for the TPH2 gene in routine practice: selection of a DNA extraction technique for PCRКурбаковК. А.KurbakovK. A.

109316, Москва, ул. Талалихина, 26

109316, Moscow, Talalikhina Str., 26

homo_ludens@vniimp.ruКибиткинаА. А.KibitkinaA. A.

109316, Москва, ул. Талалихина, 26

109316, Moscow, Talalikhina Str., 26

ankib@list.ruФедуловаЛ. В.FedilovaL. V.

к.т.н., 

109316, Москва, ул. Талалихина, 26

Cand. Sci. (Tech.), 

109316, Moscow, Talalikhina Str., 26

fedulova@vniimp.ruВасилевскаяЕ. Р.VasilevskayaE. R.

к.т.н., 

109316, Москва, ул. Талалихина, 26

Cand. Sci. (Tech.),

109316, Moscow, Talalikhina Str., 26

e.vasilevskaya@fncps.ruТолмачеваГ. С.TolmachevaG. S.

109316, Москва, ул. Талалихина, 26

109316, Moscow, Talalikhina Str., 26

tgs2991@yandex.ruФГБНУ «Федеральный научный центр пищевых систем им. В.М. Горбатова» РАНРоссияV.M. Gorbatov Federal Research Center for Food Systems of the Russian Academy of SciencesRussian Federation202123102021173E4852Copyright © Курбаков К.А., Кибиткина А.А., Федулова Л.В., Василевская Е.Р., Толмачева Г.С., 20212021Курбаков К.А., Кибиткина А.А., Федулова Л.В., Василевская Е.Р., Толмачева Г.С.Kurbakov K.A., Kibitkina A.A., Fedilova L.V., Vasilevskaya E.R., Tolmacheva G.S.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.scbmt.ru/jour/article/view/1311

Для использования биомоделей с нокаутом гена TPH2 в экспериментах ключевым моментом является разработка быстрого и высокоточного протокола генотипирования. В настоящее время генотипирование мутантных мышей проводится с помощью методов, включающих секвенирование или многоступенчатую ПЦР, являющиеся дорогостоящими и занимающими продолжительное время. В работе приведён сравнительный анализ двух методов: с помощью детекции продуктов амплификации гель-электрофореграммой и ПЦР в реальном времени. Также рассмотрена процедура выделения ДНК из фрагментов хвостов мышей. 

The development of a fast and highly accurate genotyping protocol is crucial for the use of biomodels with TPH2 gene knockout in experiments. Genotyping of mutant mice is currently carried out using methods that include sequencing or multi-stage PCR. These methods are expensive and time-consuming. This paper presents a comparative analysis of two methods: detection of amplification products by gel electrophoregram and real-time PCR. The procedure for extracting DNA from fragments of mice tails is also considered. 

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The authors declare that there are no conflicts of interest present.