224 GGACTACHVGGGTWTCTAAT V4_11R caagcagaagacggcatacgagatGTAGCCgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT V4_12R caagcagaagacggcatacgagatTACAAGgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT V4_13R caagcagaagacggcatacgagatCGTACTgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT V4_14R caagcagaagacggcatacgagatGACTGAgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT V4_15R caagcagaagacggcatacgagatGCTCAAgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT V4_16R caagcagaagacggcatacgagatTCGCTTgtgactggagttcagacgtgtgctcttccgatct GGACTACHVGGGTWTCTAAT Bold uppercase letter highlight the index sequences. Lowercase letters indicate adapter sequences necessary for flow-cell binding. Underlined lowercase letters indicate binding sites for Illumina sequencing primers. Regular uppercase letters indicate the V3 and V4 region primers (341F for the forward primer, 806R for the reverse primer). The inclusion of four maximally degenerated bases (“NNNN”) maximizes the diversity during the first four bases of the run, which is important for unique cluster identification and base-calling accuracy. Chapter 6
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