Illumina's TruSeq Sample Preparation Guide contains a table showing how to modify RNA fragmentation time to get different median insert lengths using the divalent cations and heat method in Appendix B. The median length ranges from 140 to 200 bases. Aren't all of these options wasteful if paired-end 150 bases sequencing is done? I think the ideal inset size would be 300 bases in that case, but if 0 minutes is done, the table shows 200 bases as the median size and you can't do less than 0 minutes! Is there an alternative method which doesn't result is mainly reverse-complemented pairs of reads? Our collaborators' sequencing service uses 8 minutes fragmentation and a plot of the insert sizes shows the mode of the distribution is about 120 bases and the median is about 140 bases so, the majority of reads not only are completely reverse complements of each other, but also have much (or all) of the TruSeq adapter at their ends. The reads are each 150 bases long.
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Generally smaller inserts would result in more uniform coverage of genes so for counting applications short inserts are preferred and can be sequenced only 70 cycles. For other applications such as transcriptome assembly larger fragments and high number of sequencing cycles are more efficient. Some kits such as NEBNext Ultra II enable better customisation of insert size ranging from 150bp-1kb.
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CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).
Despite this, “CRISPR helped turn genome editing from a specialized technique into...07-31-2026, 11:01 AM -
Proteomics platforms are evolving rapidly, with advances in mass spectrometry and affinity-based approaches expanding what researchers can detect and at what scale. As the field moves toward deeper proteome coverage and clinical applications, scientists face an increasingly complex landscape of tools. This article will explore how researchers are navigating these choices to find the right platform for their work.
The systematic characterization of the human proteome has...07-20-2026, 11:48 AM -
Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
...07-09-2026, 11:10 AM
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