Extracellular RNA in a single droplet of human serum reflects physiologic and disease states

We developed SILVER-seq (Small Input Liquid Volume Extracellular RNA Sequencing) to efficiently sequence both integral and fragmented cfRNAs from a small droplet (5-7 microliters) of liquid biopsy. We carried out SILVER-seq on more than 150 serum droplets from male and female donors ranging from 19 to 48 years of age. SILVER-seq detected cfRNAs from more than a quarter of the human genes, including small RNAs and fragments of mRNAs and long non-coding RNAs (lincRNAs). Overall design: 128 different samples, consisting of 32 samples from self-reported non-breast-cancer donors, 28 samples from cancer recurrence donors and 68 samples from cancer non-recurrence donors, were constructed to mols-seq libraries for cancer-normal and recurrence-nonrecurrence study.