title: rice_vitro_SHALiPE_K_replicate2
design: Total RNA was extracted from 5-day-old etiolated rice seedlings and selected by two rounds of poly(A) selection using the Poly(A) Purist MAG Kit (Ambion). 1 ?g poly(A) selected total RNA was probed using 100mM NAI at 22 C in the buffer containing 50mM Tris-HCl (pH 7.5), 0.5 mM MgCl2 and 100 mM KCl. NAI probed RNA was applied to reverse transcription with home-made RT buffer (20 mM Tris (pH 8.3), 100 mM LiCl, 3 mM MgCl2, 1 mM DTT) and random hexamers. The 3end of resultant cDNAs were ligated to a ssDNA linker using Circligase ssDNA Ligase, ligated cDNA was purified by Urea-PAGE. PCR amplification was performed using Illumina TruSeq Primers and KAPA Library Amplification Kits. PCR products were purified using agrose gel and subjected to paired-end sequencing on Illumina HiSeq 4000 platform.
library_name: RICE_KNAI_B2