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Has anyone tried using multiple staggered custom sequencing primers in order to increase diversity when amplicon sequencing? I know you can stagger primers in order to generate base diversity. Would using multiple staggered sequencing primers (ie. +1, 0, -1 base position shift) do the same trick or would it introduce a new set of problems? I'll be testing this on a miseq if that makes any difference.
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I've discussed this with coworkers and while it should be possible, it adds complexity in the setup/downstream analysis for relatively little benefit since you can also just add more phiX to offset your low diversity. You can get away with some incredibly low diversity libraries (i.e. all same sequence) using a 20% phiX spike-in.
Here's a paper that describes a similar strategy that is also cool!
https://bmcmicrobiol.biomedcentral.c...866-015-0450-4
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The introduction of single-cell sequencing has advanced the ability to study cell-to-cell heterogeneity. Its use has improved our understanding of somatic mutations1, cell lineages2, cellular diversity and regulation3, and development in multicellular organisms4. Single-cell sequencing encompasses hundreds of techniques with different approaches to studying the genomes, transcriptomes, epigenomes, and other omics of individual cells. The analysis of single-cell sequencing data i
...01-24-2023, 01:19 PM -
Single-cell sequencing is a technique used to investigate the genome, transcriptome, epigenome, and other omics of individual cells using high-throughput sequencing. This technology has provided many scientific breakthroughs and continues to be applied across many fields, including microbiology, oncology, immunology, neurobiology, precision medicine, and stem cell research.
The advancement of single-cell sequencing began in 2009 when Tang et al. investigated the single-cell transcriptomes...01-09-2023, 03:10 PM
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