In vitro selection for Microprocessor cleavage: partial-randomization in basal regions
Cleavage products (regions flanking the Drosha cleavage site) were gel purified and splint-ligated to sequencing adaptors using T4 DNA Ligase.  The 3' ligation adaptor contains a 3 nt barcode that is as the first three bases of the library.  Ligation products were reverse transcribed and amplified by for sequencing.  Libraries for four miRNA selections were mixed together before sequencing.  For each pri-miRNA, a reference pool and 1-2 selected pools (barcoded) are included.  Unique sequences that distinguish the pri-miRNAs and the individual pools are supplied below.  Indices are 0-based.    hsa-mir-125a selection:       Reference pool: CTGACC, position 67 (end 1); TATG, position 70 (end 2)       Selected pools: CTGACC, position 46 (end 1); TATGAG, position 64 (end 2)            Selected pool 1: TGA, position 0 (end 2)    hsa-mir-16-1 selection:       Reference pool: GTTGC, position 69 (end 1); GTCC, position 69 (end 2)       Selected pools: GTTGC, position 45 (end 1); GTCC, position 60 (end 2)            Selected pool 1: TGA, position 0 (end 2)            Selected pool 2: TAG, position 0 (end 2)    hsa-mir-30a selection:       Reference pool: CTTG, position 70 (end 1); AACC, position 70 (end 2)       Selected pools: CTTG, position 45 (end 1); AACC, position 60 (end 2)            Selected pool 1: CAT, position 0 (end 2)            Selected pool 2: ATG, position 0 (end 2)    hsa-mir-223 selection:       Reference pool: GAAT, position 70 (end 1); GATC, position 70 (end 2)       Selected pools: GAAT, position 48 (end 1); GATC, position 60 (end 2)            Selected pool 1: CAT, position 0 (end 2)            Selected pool 2: ATG, position 0 (end 2)