raw read of sci-ATAC-seq of sorghum leaf replicate 1 plate 4
Libraries were prepared using 24 Tn5 adapters: ME-A and 16 ME-B, each containing a 5 nt cell barcode. ME-A sequence: GGCGGTAGGCGTG NNNNN AGATGTGTATAAGAGACAG ME-B sequence: CCAACACCCGTGCG NNNNN AGATGTGTATAAGAGACAG The 24 column ME-A barcodes used were: ATACG, AGCGA, CTAAG, TGTCG, GTAGT, GTATA, ATTCC, GACTT, TTACT, CAATT, CATCT, TCGCG, TGTTT, TCCCT, GGCAG, CAAAA, GTGAG, TGGAT, CCGAG, ACTCG, GTCCT, TAAGA, ACCCC, AAACA. The 16 row ME-B barcodes used were: TCGGA, GTTTC, AGCTT, TCATT, GCTCC, ACTAA, CGAGG, ACCGG, CGGTT, GGAAC, GATGA, AACAT, CGCAC, TGACC, CGCGT, TTCCC. After Tn5 tagmentation, approximately 10 nuclei were distributed to each well of a 96-well plate. PCR was performed using 8 well-indexed forward primers (ScPCR_F) and 12 well-indexed reverse primers (scPCR_R), each containing 6 nt barcodes. ScPCR_F: ACACTCTTTCCCTACACGACGCTCTTCCGATCT NNNNNN GGCGGTAGGCGTG ScPCR_R: GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCT NNNNNN CCAACACCCGTGCG The 8 forward primer barcodes used were: GATCAG, TAGCTT, GGCTAC, CTTGTA, AGTCAA, AGTTCC, ATGTCA, CCGTCC. The 12 reverse primer barcodes used were: GTAGAG, GTCCGC, GTGAAA, GTGGCC, GTTTCG, CGTACG, GAGTGG, GGTAGC, ACTGAT, ATGAGC, ATTCCT, CAAAAG. After sequencing, the data from each plate was demultiplexed using the 24x16 cell and 8x12 well barcodes. Additionally, a 6 nt plate barcode was added to the cell+well barcode before combining the data from each plate.