title: CLICK-seq of DL1 cells following Brat RNAi
design: 5 ug total RNA was subjected to poly(A) mRNA isolation.  We reverse transcribed 10 uL of eluted RNA with Genomic Adaptor_6N and azido-nucleotide (AzVTPs) / dNTPs (35:1 ratio). We click-ligated the UMI-click-adapter to the 5 end of the cDNA. We then amplified the cDNA for 15 cycles with 3'-phosphorothioate modified i5 and i7 indexing primers, and purified and size-selected with Speedbeads. Libraries were pooled and sequenced single-end, 100 nt reads on a NovaSeq 6000 NGS