Illumina HiSeq 2000 sequencing; Extensive small RNAs sequestration in diverse eukaryotes
Extensive small RNAs sequestration in diverse eukaryotes
Total RNA was extracted from N. benthamiana tissue based on the White and Kaper protocol (/White, J.L., and Kaper, J.M. (1989). A simple method for detection of viral satellite RNAs in small plant tissue samples. J. Virol. Methods 23, 83 to 93). Total RNA from the all other organisms (A. thaliana, M. musculus, C. elegans, D. melanogaster) were extracted by TRIzol reagent (Life Technolgies). To make the small RNA libraries, this total RNA was used directly for Illumina TruSeq small RNA cloning protocol (RS-200-9002DOC) (NSS= non size selected). Or was subject to this protocol after small RNA size selection, by extraction of the 18-30nt band from a denaturing (formamide) polyacrylamide gel (FSS) or after small RNA size selection by extraction of the 18-30nt band from a fully denaturing formaldehyde polyacrylamide gel (FDFSS - described in detail by Harris et al,. in prep).
Experimental Factor: protocol
NSS
Experimental Factor: organism
Arabidopsis thaliana