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• What will be the minimum number of biological replicates (plant samples) and how many biological replicates are needed for RNA-Seq using HIseq ?
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Depends on your experimental design, but absolutely not less than two per experimental condition. It doesn't matter which platform you use. -
One more question about the "replicate" definition; if you have a cDNA from a biological sample (for a RNA-seq experiment) and you use it into two PCR reactions with different barcodes (like the one you do before the Illumina run), the barcode1 and barcode2 samples from the same cDNA can be considered as "replicates".
Suppose you want statistical power in your data, and you want to test differential gene expression in programs like DESeq (where it wants replicates to check the so called "shot-noise"); in this case, can they be considered good "replicates"?
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These are more like technical replicates to me...Originally posted by cascoamarillo View PostComment
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Yes, these replicates are only testing the reproducibility of the sequencing protocol, which is extremely high. Biological reproducibility is what we're interested in. I would just pool the data from technical replicates as a single biological replicate.Comment
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I would do no less than three independent biological replicates (i.e. RNA isolated from separate biological samples) of each condition. You can get meaningful results from 2 biological reps, but you are really restricting yourself by doing this, particularly on a HiSeq where you can easily multiplex and still get a lot of reads per sample even on a single lane.Comment
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Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
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With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.
Introduction
Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...05-22-2026, 06:42 AM
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