Extraction of Total DNA and RNA from Marine Filter Samples and Generation of a cDNA as Universal Template for Marker Gene Studies

Methods Mol Biol. 2017:1539:13-22. doi: 10.1007/978-1-4939-6691-2_2.

Abstract

Microbial communities play an important role in marine ecosystem processes. Although the number of studies targeting marker genes such as the 16S rRNA gene has been increased in the last few years, the vast majority of marine diversity is rather unexplored. Moreover, most studies focused on the entire bacterial community and thus disregarded active microbial community players. Here, we describe a detailed protocol for the simultaneous extraction of DNA and RNA from marine water samples and for the generation of cDNA from the isolated RNA which can be used as a universal template in various marker gene studies.

Keywords: Marker gene studies; Metagenomics; Metatranscriptomics; Microbial diversity; Microbial functions.

MeSH terms

  • Aquatic Organisms / genetics*
  • Biodiversity
  • DNA*
  • DNA, Complementary*
  • Genetic Markers
  • Marine Biology / methods
  • Metagenome*
  • Metagenomics* / methods
  • RNA*

Substances

  • DNA, Complementary
  • Genetic Markers
  • RNA
  • DNA