A specific combination of dual index adaptors decreases the sensitivity of amplicon sequencing with the Illumina platform.

ABSTRACT: Amplicon sequencing is a powerful approach in microbiome studies as it detects live organisms with high sensitivity. This approach determines the composition of sequence variants of marker genes using high-throughput DNA sequencers. The use of dual index adaptors is the fundamental technique for pooling DNA libraries for Illumina sequencers and is believed not to affect the results. However, here, we observed a decrease of sequence quality in samples containing a specific combination of indexes, namely N704 and S507 in Nextera kits, in multiple runs on the Illumina MiSeq sequencer operated in different facilities. This decrease was also observed when sequencing randomly fragmented DNA of Escherichia coli and was not observed when either individual adaptor was used. Each end of the DNA libr

SUBMITTER: Hirose Y 

PROVIDER: S-EPMC7547650 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature