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Usually, the sacrifice of snakes is necessary for venom gland extraction to identify protein-coding transcripts; however, the venom can be used as a source of transcripts. Although there are methods for obtaining RNA from venom, no comparative analysis has been conducted in the Bothrops genus. In the present study, we compared four commercial methods for RNA purification and cDNA synthesis from venom (liquid, lyophilized, or long-term storage) of four clinically relevant species of Peruvian Bothrops. Our results show that the TRIzol method presents the highest yield of RNA purified from venom (59 \u00b1 11 ng\/100 \u00b5L or 10 mg). The SuperScript First-Strand Synthesis System kit produced high amounts of cDNA (3.2 \u00b1 1.2 ng cDNA\/ng RNA), and the highest value was from combination with the Dynabeads mRNA DIRECT kit (4.8 \u00b1 2.0 ng cDNA\/ng RNA). The utility of cDNA was demonstrated with the amplification of six relevant toxins: thrombin-like enzymes, P-I and P-III metalloproteinases, acid and basic phospholipases A2, and disintegrins. To our knowledge, this is the first comparative study of RNA purification and cDNA synthesis methodologies from Bothrops genus venom.<\/jats:p>","DOI":"10.3390\/ijms241311161","type":"journal-article","created":{"date-parts":[[2023,7,7]],"date-time":"2023-07-07T01:41:36Z","timestamp":1688694096000},"page":"11161","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Comparison of Four Methods of RNA Extraction and cDNA Synthesis from The Venom of Peruvian Snakes of the Genus Bothrops of Clinical Importance"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9385-6921","authenticated-orcid":false,"given":"Daniel","family":"Torrej\u00f3n","sequence":"first","affiliation":[{"name":"Laboratorio de Biolog\u00eda Molecular, Facultad de Ciencias Biol\u00f3gicas, Universidad Nacional Mayor de San Marcos, Av. Venezuela Cdra 34 S\/N, Ciudad Universitaria, Lima Cercado, Lima 15081, Peru"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6112-2584","authenticated-orcid":false,"given":"Javier","family":"C\u00e1rdenas","sequence":"additional","affiliation":[{"name":"Laboratorio de Bioqu\u00edmica, Facultad de Ciencias de la Salud, Universidad Nacional del del Callao, Av. Juan Pablo \u0399\u0399 306, Bellavista 07011, Peru"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diana","family":"Ju\u00e1rez","sequence":"additional","affiliation":[{"name":"Laboratorio de Gen\u00f3mica, Pontificia Universidad Cat\u00f3lica del Per\u00fa, Av. 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Venezuela Cdra 34 S\/N, Ciudad Universitaria, Lima Cercado, Lima 15081, Peru"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7297-0263","authenticated-orcid":false,"given":"Mariana","family":"Legu\u00eda","sequence":"additional","affiliation":[{"name":"Laboratorio de Gen\u00f3mica, Pontificia Universidad Cat\u00f3lica del Per\u00fa, Av. Universitaria 1801, Campus Principal, San Miguel 15088, Peru"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8582-4399","authenticated-orcid":false,"given":"F\u00e9lix A.","family":"Urra","sequence":"additional","affiliation":[{"name":"Laboratorio de Plasticidad Metab\u00f3lica y Bioenerg\u00e9tica, Programa de Farmacolog\u00eda Cl\u00ednica y Molecular, Instituto de Ciencias Biom\u00e9dicas, Facultad de Medicina, Universidad de Chile, Santiago 8380453, Chile"},{"name":"Network for Snake Venom Research and Drug Discovery, Av. Independencia 1027, Santiago 7810000, Chile"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eladio F.","family":"S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Network for Snake Venom Research and Drug Discovery, Av. 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