Subtractive Hybridization

Sample Processing and cDNA Preparation for Microbial Metatranscriptomics in Complex Soil Communities

“…This technique involves the subtraction of a specific set of nucleic acids (in this case rRNA) from a reference sample (total RNA) using probes with Combination of subtractive hybridization and exonuclease treatment Mettel et al (2010), He et al (2010) Size separation by gel electrophoresis McGrath et al (2008) sequences that are complementary to the set to be subtracted (rRNA) (Pang et al, 2004;Su & Sordillo, 1998;Winstanley, 1998). rRNA can be partially subtracted by the commercial kits MICROBExpress Bacterial mRNA Enrichment (Ambion, Austin, TX, USA) and Ribo-Zero Magnetic kit (Bacteria) (Epicentre, Madison, WI, USA) through capture 16S and 23S binding oligonucleotides immobilized on magnetic microbeads.…”