45 research outputs found
SemiQuant/RNAseq_pipeline: RNAseq_pipeline_v1.0.0
No full textIllumina RNA sequencing processing pipeline and post processing script
A simple, single-tube overlapping amplicon-targeted Illumina sequencing assay.
No full textTargeted amplicon sequencing to identify pathogens, resistance-conferring mutations, and strain types is an important tool in diagnosing and treating infections. However, due to the short read limitations of Illumina sequencing, many applications require the splitting of limited clinical samples between two reactions. Here, we outline hairpin Illumina single-tube sequencing PCR (hissPCR) which allows for the generation of overlapping amplicons containing Illumina indexes and adapters in a single tube, effectively extending the Illumina read length while maintaining reagent and sample input requirements
The protocol in PDF format available from protocols.io is provided as supporting information File 1, with the caption: S1: Step-by-step protocol, also available on protocols.io.
No full textSequence data is available on SRA under the accession number PRJNA949687. (PDF)</p
Expected results.
No full textA) hissPCR generated amplicons using two forward and two reverse overlapping primers in a single tube. Amplicons contain either no Illumina adapter, one Illumina adapter, or two Illumina adapters per amplicon. B) The relative proportion of amplicons in the sample containing both Illumina adapters. C) Melt curves showing the expected amplicons for each primer set and the ratio of amplicons containing both Illumina adapters.</p
