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.

ABSTRACT:

Background

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 reduces the overall number of sequencing reads for data analysis. To overcome such low diversity issues during amplicon sequencing on illumina platforms, we developed a high throughput single amplicon sequencing method by introducing 'N' (0-10) spacers in target gene amplification primers that are pooled for simple handling.

Result

We evaluated the efficiency of 'N' (0-10) spacer-linked primers by targeting bacterial 16S V3-V4 region, demonstrating heterogeneous base libra

SUBMITTER: Naik T 

PROVIDER: S-EPMC10037784 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature