ABSTRACT
The analysis of gene expression in soil microbiomes via a suite of methods is subjected to an array of considerable limitations. Microarray hybridization analysis has been considered to represent a major advance in the semiquantitative analysis of a large number of genes in parallel, which could be also useful for RNA-based studies to assess activity pattern. The most commonly used enzymes for cDNA synthesis from RNA extracted from soil are avian myeloblastosis virus and Moloney murine leukemia virus reverse transcriptases. The coupling of retro-transcribing polymerase chain reaction with fluorescence techniques and advanced technology capable of automated detection and quantification of mRNAs has led to the development of new technologies that have dramatically changed gene expression studies. Metatranscriptomics allows the nontargeted analysis of transcripts produced in a complex microbial community at any point in time and space. mRNA-based analyses have the potential to elucidate the resilience of soil microbiomes to perturbations, such as pollutant input and climate change.
