Rna clean kit

The reaction was carried out in 50 μL of solution containing 100 mM Tris⋅HCl, pH 8.0, 10 mM iodoacetic acid, 500 ng 38-nt NAD-RNAs, or dpCoA-RNAs and 50 units of murine RNase inhibitor (NEB) at 37 °C for 15 min. The RNAs were purified with RNA clean kit (Zymo) following the manufacturer’s instructions.

Samples were drawn to extract RNA for microarray analysis such that the product of volume and OD730nm was 50.0. The same RNA samples were also used for Real-time PCR analysis for validation of microarray results [18 (link)]. Nine samples were drawn from two consecutive cycles under LL (S1 Fig). Microarray analysis was performed at Mogene (St. Louis, USA) as described earlier [19 (link)]. Briefly, total RNA from individual samples and an equi-molar mixture of all samples as control were labeled with Cy3 and Cy5, respectively. Unbound dyes were removed using RNA clean kit (Zymo Research, Irvine, CA). Two biological replicates, two technical replicates and a dye swap were analyzed for each sample from individual time points. The microarray dataset submitted to Gene Expression Omnibus (GEO) database, accession no. GSE65429.

Size selection was conducted using the RNA Clean kit (Zymo) according to the manufacturer’s instructions. The detailed method is described in SI Appendix, Supplementary Methods.