دورية أكاديمية
Convective polymerase chain reaction in standard microtubes.
| العنوان: | Convective polymerase chain reaction in standard microtubes. |
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| المؤلفون: | Garafutdinov RR; Institute of Biochemistry and Genetics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: garafutdinovr@gmail.com., Chemeris DA; Institute of Biochemistry and Genetics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: chemeris@gmail.com., Sakhabutdinova AR; Institute of Biochemistry and Genetics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: sakhabutdinova.a.r@gmail.ru., Moiseev KV; Mavlyutov Institute of Mechanics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: constgo@anrb.ru., Urmancheev SF; Mavlyutov Institute of Mechanics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: said52@mail.ru., Mikhaylenko CI; Mavlyutov Institute of Mechanics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: const@uimech.org., Privalov LY; Mavlyutov Institute of Mechanics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: lavrenty.privalov@gmail.com., Chemeris AV; Institute of Biochemistry and Genetics, Ufa Federal Research Center, Russian Academy of Sciences, 450054, Prosp. Oktyabrya, 71, Ufa, Bashkortostan, Russia. Electronic address: chemeris@anrb.ru. |
| المصدر: | Analytical biochemistry [Anal Biochem] 2022 Mar 15; Vol. 641, pp. 114565. Date of Electronic Publication: 2022 Jan 22. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: United States NLM ID: 0370535 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1096-0309 (Electronic) Linking ISSN: 00032697 NLM ISO Abbreviation: Anal Biochem Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: <2000- > : San Diego, CA : Elsevier Original Publication: Orlando Fl : Academic Press |
| مواضيع طبية MeSH: | COVID-19/*diagnosis , COVID-19 Nucleic Acid Testing/*instrumentation , Polymerase Chain Reaction/*instrumentation , SARS-CoV-2/*isolation & purification, COVID-19/virology ; COVID-19 Nucleic Acid Testing/economics ; COVID-19 Nucleic Acid Testing/methods ; Convection ; Humans ; Polymerase Chain Reaction/economics ; Polymerase Chain Reaction/methods ; RNA, Viral/analysis ; RNA, Viral/genetics ; SARS-CoV-2/genetics ; Temperature ; Time Factors |
| مستخلص: | Polymerase chain reaction (PCR) is the most widely used method for nucleic acids amplification. To date, a huge number of versatile PCR techniques have been developed. One of the relevant goals is to shorten PCR duration, which can be achieved in several ways. Here, we report on the results regarding nucleic acids amplification by convective PCR (cPCR) in standard 0.2 ml polypropylene microtubes. The following conditions were found to be optimal for such amplification: 1) 70 μl reaction volume, 2) the supply of external temperature 145°С for the denaturation zone and 0°С for the annealing zone, 3) ∼30° inclination of the microtube main axis, 4) the use of nearby primers, and 5) duration of the reaction 15-20 min. At these conditions, the amplification products are accumulated in an amount sufficient to be registered by gel electrophoresis, and high sensitivity of the reaction comparable to that of conventional PCR is achieved. cPCR provided the reliable detection of SARS-CoV-2 coronavirus RNA isolated from nasopharyngeal swabs of COVID-19 patients. (Copyright © 2022 Elsevier Inc. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Convection; Nearby primers; Polymerase chain reaction (PCR); SARS-CoV-2 |
| المشرفين على المادة: | 0 (RNA, Viral) |
| تواريخ الأحداث: | Date Created: 20220125 Date Completed: 20220215 Latest Revision: 20220215 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1016/j.ab.2022.114565 |
| PMID: | 35074320 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1096-0309 |
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| DOI: | 10.1016/j.ab.2022.114565 |