I am doing RNA-seq using illumina kit. But i am confusing on the insert size. When performing gel seclection after cDNA being ligated with adaptor, the 200 bp size with cDNA and adaptor is seclected on gel. Because the adaptor length on the two end of cDNA is 60 bp, the cDNA should be 140 bp. After performing PCR using 200 bp size (adaptor plus cDNA) as template, the final product should be 260 bp. Because the each PCR primer has 30 nt overhang compared with template, in theory, the final size should be 200 + 60 (sense and antisense primer), 260 bp. However, the factual size is still around 200 bp by Agilent bioanalyzer from illumina protocol. How to explain the difference between theoretical 260 bp and realistic 200 bp? What's wrong with the agilent analysis or gel seclection?
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The immune system’s power comes from its genetic diversity, allowing myriad threats to be neutralized through first recognizing foreign antigens. That diversity is also what makes the immune system so difficult to study. Recent advances in sequencing technology and computational biology, however, are giving researchers new tools to understand immune responses and immune-related diseases in greater detail.
This convergence of genetics, immunology, and computation...09-01-2026, 05:41 AM