MethodsX (Jan 2019)

Evaluation of two independent protocols for the extraction of DNA and RNA from different tissues of sea cucumber Isostichopus badionotus

  • Carlos Puch-Hau,
  • Itzel A. Sánchez-Tapia,
  • Victoria Patiño-Suárez,
  • Miguel A. Olvera-Novoa,
  • Mariel Gullian Klanian,
  • Mercedes Quintanilla-Mena,
  • Omar Zapata-Pérez

Journal volume & issue
Vol. 6
pp. 1627 – 1634

Abstract

Read online

Isostichopus badionotus is a sea cucumber species of great ecological and economic relevance for Mexico and Central American and Caribbean countries; however, the protocols for the extraction of the nucleic acids have not yet been published. In this study, we describe the first protocols to obtain DNA and RNA from different tissues of I. badionotus, which include the respiratory tree, gonad, longitudinal muscle bands, anterior intestine and cloaca. The extraction of high-quality DNA was performed using the DNeasy Blood & Tissue kit (Qiagen, Valencia, CA, USA) with minor modifications in different points of the protocol.Concerning the RNA, the method of TRIzol was used. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous RNases or when the separation of cytoplasmic RNA from nuclear RNA is impractical. The methodologies used in this study allowed us to obtain DNA and RNA of high quality and integrity in the different tissues of I. badionotus, which will be the basis for future genomic and transcriptomic studies. • The successful extraction of DNA and RNA was achieved in the different tissues of I. badionotus. • The concentrations of DNA and RNA obtained were adequate for a diversity of analyses at a molecular level. Method name: I. badionotus DNA and RNA isolation, Keywords: Holothuroid, Molecular biology, Genetics, Nucleic acid