
Задачи иÑÑледованиÑ: - Разработать ÑоÑтав лизирующего буфера, позволÑющего Ñффективно ÑкÑтрагировать генетичеÑкий материал из образца; - Разработать протокол термичеÑкой ÑкÑтракции генетичеÑкого материала из образца; - Оценить ÑффективноÑть различных ÑпоÑобов ÑкÑтракции генетичеÑкого материала и выбрать оптимальный. Объектом иÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ð´Ð°Ð½Ð½Ð¾Ð¹ работы ÑвлÑÑŽÑ‚ÑÑ Ñ€Ð°Ð·Ð»Ð¸Ñ‡Ð½Ñ‹Ðµ методы ÑкÑтракции генетичеÑкого материала из образца. Предметом иÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ ÑвлÑетÑÑ Ð¸Ð·ÑƒÑ‡ÐµÐ½Ð¸Ðµ наиболее проÑтого и хорошо подходÑщего Ð´Ð»Ñ Ð¸ÑÐ¿Ð¾Ð»ÑŒÐ·Ð¾Ð²Ð°Ð½Ð¸Ñ Ð² реакции изотермичеÑкой амплификации ÑпоÑоба ÑкÑтракции генетичеÑкого материала из образца. При напиÑании литературного обзора иÑпользовалиÑÑŒ Ñледующие базы данных: eLIBRARY.RU, PubMed. СтатиÑтичеÑкую обработку ÑкÑпериментальных данных оÑущеÑтвлÑли Ñ Ð¸Ñпользованием пакета программ Microsoft Excel. Работа была выполнена на базе лаборатории ÐИК «Ðанобиотехнологии» Санкт ПетербургÑкого гоÑударÑтвенного политехничеÑкого универÑитета Петра Великого. Ð’ ходе иÑÑÐ»ÐµÐ´Ð¾Ð²Ð°Ð½Ð¸Ñ Ñтало ÑÑно, что наиболее Ñффективным методом ÑкÑтракции генетичеÑкого материала из образца ÑвлÑетÑÑ Ð¼ÐµÑ‚Ð¾Ð´ Ñ Ð²Ð¾Ð´Ð¾Ð¹ Milli-Q. Ð’Ñе протеÑтированные ÑоÑтавы буферов показали результаты хуже, чем метод термичеÑкой ÑкÑтракции водой Milli-Q. Был ÑоÑтавлен протокол ÑкÑтракции.Â
The tasks of the study: - To develop the composition of a lysing buffer that allows efficient extraction of genetic material from a sample; - To develop a protocol for thermal extraction of genetic material from a sample; - Evaluate the effectiveness of various methods of extracting genetic material and choose the best one. The object of the study of this work is various methods of extraction of genetic material from a sample. The subject of the study is the study of the simplest and most suitable method for extraction of genetic material from a sample for use in an isothermal amplification reaction. When writing the literary review, the following databases were used: eLibrary.RU, PubMed. Statistical processing of experimental data was carried out using the Microsoft Excel software package. The work was carried out in the analytical center of nano- and biotechnologies of Peter the Great St. Petersburg Polytechnic University. During the study, it became clear that the most effective method of extracting genetic material from a sample is the Milli-Q water extraction method. All tested buffer formulations showed worse results than the Milli-Q water extraction method. An extraction protocol was drawn up.
ÑкÑÑÑакÑиÑ, ÑкÑпÑеÑÑ-диагноÑÑика, амплиÑикаÑиÑ, extraction, CRISPR-CAS, amplification, express diagnostics
ÑкÑÑÑакÑиÑ, ÑкÑпÑеÑÑ-диагноÑÑика, амплиÑикаÑиÑ, extraction, CRISPR-CAS, amplification, express diagnostics
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