Single-cell whole-genome sequencing (WGS) is critical for characterizing dynamic intercellular changes in DNA. Current sample preparation technologies for single-cell WGS are complex, expensive, and suffer from high amplification bias and errors. Here, we describe Digital-WGS, a sample preparation platform that streamlines high-performance single-cell WGS with automatic processing based on digital microfluidics. Using the method, we provide high single-cell capture efficiency for any amount and types of cells by a wetted hydrodynamic structure. The digital control of droplets in a closed hydrophobic interface enables the complete removal of exogenous DNA, sufficient cell lysis, and lossless amplicon recovery, achieving the low coefficient of variation and high coverage at multiple scales. The single-cell genomic variations profiling performs the excellent detection of copy number variants with the smallest bin of 150 kb and single-nucleotide variants with allele dropout rate of 5.2%, holding great promise for broader applications of single-cell genomics.
Biomarkers
based on DNA methylation have attracted wide attention
in biomedical research due to their potential clinical value. Therefore,
a sensitive and accurate method for DNA methylation detection is highly
desirable for the discovery and diagnostics of human diseases, especially
cancers. Here, an integrated, low-cost, and portable point-of-care
(POC) device is presented to analyze DNA methylation, which integrates
the process of pyrosequencing in a digital microfluidic chip. Without
additional equipment and complicated operation, droplets are manipulated
by patterned electrodes with individually programmed control. The
system exhibited an excellent sensitivity with a limit of detection
(LOD) of 10 pg and a comparable checkout down to 5% methylation level
within 30 min, which offered a potential substitute for the detection
of DNA methylation. With the advantages of portability, ease of use,
high accuracy, and low cost, the POC platform shows great potential
for the analysis of tumor-specific circulating DNA.
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