| Chapter title |
Metatranscriptomics of Soil Eukaryotic Communities
|
|---|---|
| Chapter number | 16 |
| Book title |
Microbial Environmental Genomics (MEG)
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3369-3_16 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3367-9, 978-1-4939-3369-3
|
| Authors |
Rajiv K. Yadav, Claudia Bragalini, Laurence Fraissinet-Tachet, Roland Marmeisse, Patricia Luis |
| Editors |
Francis Martin, Stephane Uroz |
| Abstract |
Functions expressed by eukaryotic organisms in soil can be specifically studied by analyzing the pool of eukaryotic-specific polyadenylated mRNA directly extracted from environmental samples. In this chapter, we describe two alternative protocols for the extraction of high-quality RNA from soil samples. Total soil RNA or mRNA can be converted to cDNA for direct high-throughput sequencing. Polyadenylated mRNA-derived full-length cDNAs can also be cloned in expression plasmid vectors to constitute soil cDNA libraries, which can be subsequently screened for functional gene categories. Alternatively, the diversity of specific gene families can also be explored following cDNA sequence capture using exploratory oligonucleotide probes. |
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