GSM8582123: B73_premeiotic_anther_sRNA_rep2; Zea mays; ncRNA-Seq
Total RNA was isolated from embryos, anthers and male germ cells using TRIzol (Takara, 9109). MgCl2 (final concentration: 0.2 M) was added to prevent loss of sRNAs with low GC content (Kim et al., 2012). NaAc (pH 5.2) and glycogen were added to promote RNA precipitation. The extracted RNA was stored at −80°C in LoBind tubes (Eppendorf, 022431021). Libraries for small RNA sequencing were constructed as previously described (Yang et al., 2016). In brief, total RNA was ligated to 3′ adaptor with adenylation using T4 RNA ligase 2, truncated KQ (NEB, M0373L). Following annealing with reverse transcription primer, RNA was ligated to 5′ RNA adaptor using T4 RNA ligase 1 (NEB, M0204L). After reverse transcription and PCR amplification, 135–160-bp products were gel-purified and sent for sequencing. Libraries were pair-end sequenced with Illumina Nova platform by Annoroad Gene Technology (Beijing). sRNA-seq
GEO
GSM8582123_r1