Adult control and postdauer animals were collected as previously described (Hall, Beverly, et al 2010). Dauer animals were collected 24 hours after SDS treatment, following the postdauer growth strategy. L3 animals were grown according to the control adult strategy, but collected at the L3 stage.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted with Trizol reagent (Sigma) from flash-frozen staged worms, and 25 ug was subjected to 5'-triphosphate removal treatment with Tobacco Acid Phosphatase (TAP; Epicentre). This treatment left a 5’-monophosphate on siRNAs. Small RNAs between 18-32 nt were size selected and subjected to linker ligation reactions and amplification as described previously (Lau et al 2001 Science). Two biological replicates were sequenced for adult libraries, one biological replicate for L3 larvae and dauer libraries. sRNA libraries were sequenced on an Illumina GAII machine, yielding between 4 and 14 million reads per library that matched to the C. elegans genome.
Library strategy
RNA-Seq
Library source
transcriptomic
Library selection
cDNA
Instrument model
Illumina Genome Analyzer II
Description
20110310_8_Dauer
Data processing
Data tables include the reads and counts after barcode removal and trimming of adaptor sequences.
