High-quality single amplicon sequencing method for illumina MiSeq platform using pool of ‘N’ (0–10) spacer-linked target specific primers without PhiX spike-in

• Published in: BMC genomics Vol. 24; no. 1; pp. 141 - 11
• Main Authors: Naik, Tejali, Sharda, Mohak, C P, Lakshminarayanan, Virbhadra, Kumar, Pandit, Awadhesh
• Format: Journal Article
• Language: English
• Published: England BioMed Central 23.03.2023; BMC
• Subjects: Amplicon sequencing; Bacteria - genetics; Base composition; Data analysis; E coli; Gene amplification; Gene Library; Genetic engineering; Genomics; High-Throughput Nucleotide Sequencing - methods; Illumina; Lasers; Libraries; Microbiota - genetics; Microorganisms; PhiX spike-in; Primers; RNA, Ribosomal, 16S - genetics; Sequence Analysis, DNA - methods; Software; Spacer; Spacers; Taxonomy; ‘N’ (0–10) spacer-linked primer pool; ‘N’ (0–10) spacer-linked primer pool; PhiX spike-in; Illumina; Amplicon sequencing

Illumina sequencing platform requires base diversity in the initial 11 cycles for efficient cluster identification and colour matrix estimation. This limitation yields low-quality data for amplicon libraries having homogeneous base composition. Spike-in of PhiX library ensures base diversity but red...