Single Read Sequencing
Single Read Sequencing - Related Videos
08:57
Amplicon Sequencing using the Long-Read Sequencing Technologies
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2025
This protocol was optimized for targeted deep sequencing of 18 drug-resistance regions in Mycobacterium tuberculosis using a long-read sequencing platform, followed by analysis with a tuberculosis-specific bioinformatics pipeline designed for long-read data.
06:56
Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing
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Cited by 17 •
2017
Described here is a method for the extraction, purification, and quality control of genomic DNA from the obligate biotrophic fungal pathogen, powdery mildew, for use in long-read genome sequencing.
14:49
Single Read and Paired End mRNA-Seq Illumina Libraries from 10 Nanograms Total RNA
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Cited by 30 •
2011
Here we describe a method for preparation of both single read and paired end Illumina mRNA-Seq sequencing libraries for gene expression analysis based on T7 linear RNA amplification. This protocol requires only 10 nanograms of starting total RNA and generates highly consistent libraries representing whole transcripts.
10:34
Ultra-long Read Sequencing for Whole Genomic DNA Analysis
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Cited by 28 •
2019
Long-read sequences greatly facilitate the assembly of complex genomes and characterization of structural variation. We describe a method to generate ultra-long sequences by nanopore-based sequencing platforms. The approach adopts an optimized DNA extraction followed by modified library preparations to generate hundreds of kilobase reads with moderate coverage from human cells.
09:45
Detection of Copy Number Alterations Using Single Cell Sequencing
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Cited by 9 •
2017
Single cell sequencing is an increasingly popular and accessible tool for addressing genomic changes at high resolution. We provide a protocol that uses single cell sequencing to identify copy number alterations in single cells.
FAQs
After adapter ligation, library fragments are immobilized so the sequencing system can follow each fragment during base-by-base copying or reading. In sequencing by synthesis, the newly determined bases form a continuous read from one fragment end. This arrangement links the resulting sequence directly to the original library fragment, supporting downstream identification or measurement of genetic material.
The main information trade-off is positional context. Because only one end of each fragment is read, the result generally provides less positional information than paired-end sequencing, which supplies more positional information. Single read sequencing can therefore be attractive when straightforward coverage is more important than added positional context.
Fragment length and genome complexity are important suitability factors for Single Read Sequencing. Applications are most appropriate when these characteristics fit the intended analysis, because a single-end read provides less positional information than paired-end data. Considering both variables helps researchers judge whether gene identification, transcript quantification, variant detection, or sample characterization is the strongest use.
Sequence reads can be used to identify genes and quantify transcripts, making the approach relevant to both genomic and transcriptomic studies. Its value is not limited to cataloging sequence: read counts or sequence matches can support measurement of genetic material, provided fragment length and genome complexity are suitable for the biological sample and the study's intended analysis.
It is especially useful when a study needs efficient, cost-effective coverage rather than the additional positional information associated with paired-end sequencing. This makes it a practical option across genomics, transcriptomics, and molecular biology, assuming the library's fragment length and the sample's genome complexity support the intended profiling task.
Single read sequencing can characterize microbial and environmental samples in addition to genomic or transcriptomic material. The resulting profiles may reveal genes, support transcript quantification, or detect small variants, depending on the sample and the suitability of its fragment length and genome complexity. This flexibility makes it useful for molecular profiling and sample characterization in biology.