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Hasby, F.

Publications and source records attributed to Hasby, F..

2 recordsLinked to original sources
bioRxiv2025

Tech Note - Adaptive Sampling: targeted Oxford Nanopore long-read sequencing

Oxford Nanopore Technology (ONT) has developed a long-read sequencing method known as "adaptive" or "Read Until" sequencing. Unlike Illumina sequencing, this ONT read data is available in real-time during the run, rather than only being available when the sequencing run is complete and it focuses on a specified region of interest (ROI) within the sequence data. We investigated how this technique compared to standard sequencing under a variety of conditions and found it consistently gave a multifold increase in coverage of the ROI compared to standard sequencing, though a strong positional effect was observed for ROI efficacy. Adaptive sampling can be used to selectively enrich or deplete sequences from a library in real-time, enhancing coverage of regions of interest without requiring additional library pre-processing

genomics↗
bioRxiv2025

Adapter dilution and input optimisation for Qiagen QIAseq miRNA Library kit

MicroRNAs (miRNAs) are small, non-coding RNA that play a critical role in regulating gene expressions that are important for a multitude of biological processes. In the preparation of miRNA sequencing libraries by using QIAseq miRNA library kit, adapter dimers might occur inadvertently and compromise the sequencing performance. In this technical note, we tested different adapter dilution strategies to minimise or diminish adapter dimers without compromising data quality. In our experiments, we found that RNA input amount plays a larger role in adapter dimer formation rather than the availability of the adapters itself. We suggest preparing the sequencing library with 10 times more RNA than the recommended input amount of the chosen protocol, e.g. following the adapter dilution and PCR cycle number of the standard 1 ng protocol with 10 ng RNA input. We also suggest analysing the data with deduplication by using unique molecular identifiers (UMIs) as it helps remove unusable reads.

genomics↗