IsoSeq sequencing of transcriptome of Heterodera trifolii
SMRTLink 6.0 Sequel IsoSeq3 classify Separates Reads Of Insert reads into full-length and non-full-length based on primer and (optionally) polyA tail detection. The output sequences are stripped of primers and polyA tails and re-oriented. Each output sequence represents exactly one read from a single ZMW (single molecule) and accuracy for each read varies depending on raw read length and transcript length, which may be anywhere between 85-100%.SMRTLink 6.0 Sequel IsoSeq ICE and Arrow on cluster Time and memory-intensive isoform-level clustering algorithm that outputs high-quality consensus transcript sequences. Every output sequence is an isoform consensus that is the consensus of many different reads that are considered by the algorithm to be the same isoform. Arrow is used to generate the consensus and the associated consensus QVs are used to estimate how good the final consensus accuracy is. Based on the Arrow estimated accuracy and user parameter (Default: 99% accurate), the consensus sequences are categorized as HQ (high-quality, above 99% or user-defined cutoff) and LQ (low-quality).