Phased nucleotide inserts for sequencing low-diversity RNA samples from in vitro selection experiments.

ABSTRACT: In vitro selection combined with high-throughput sequencing is a powerful experimental approach with broad application in the engineering and characterization of RNA molecules. Diverse pools of starting sequences used for selection are often flanked by fixed sequences used as primer binding sites. These low diversity regions often lead to data loss from complications with Illumina image processing algorithms. A common method to alleviate this problem is the addition of fragmented bacteriophage PhiX genome, which improves sequence quality but sacrifices a portion of usable sequencing reads. An alternative approach is to insert nucleotides of variable length and composition ("phased inserts") at the beginning of each molecule when adding sequencing adaptors. This approach preserves read dept

SUBMITTER: Bendixsen DP 

PROVIDER: S-EPMC7373987 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature