Zea mays B73, primary and seminal root tip (0-1mm), Nuclear EU nascent RNA-seq, biorep3
Biotinylated RNA was incubated with streptavidin beads using the Invitrogen Click-it Nascent RNA capture kit to enrich for EU-RNAs. First strand cDNA synthesis was performed with Superscript IV First Strand Synthesis System using random hexamer priming. Libraries were made using a NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (starting at step 1.4.1 because cDNAs were previously made) in combination with NEBNext Multiplex Oligos for Illumina, Unique Dual Index UMI Adaptors RNA Set 1. The protocol to target 200 nt inserts was followed and the adapter ligated libraries were amplified for 14 PCR cycles.