How to Sequence DNA from Tiny Samples Using Specialized Plates

A method for preparing DNA for genetic sequencing by splitting tiny samples into a 5184-well plate to ensure accurate data from very few cells.

What it covers

This patent describes a specific workflow for preparing DNA for high-throughput sequencing, designed to handle samples with as few as 10 to 500 cells. The process involves distributing single-stranded DNA into a 5184-well plate, where the volume of each well is strictly controlled to 350 nL. By keeping the sample size small and the distribution precise, the method ensures that homologous chromosome fragments from both parents are unlikely to end up in the same well. After amplification and fragmentation using a transposase enzyme, the resulting DNA library is ready for sequencing.

What it doesn't cover

• —Does not cover sequencing methods that do not use the specific 5184-well plate configuration.
• —Does not cover general whole genome amplification techniques performed outside of this specific plate-based workflow.
• —Does not cover sequencing libraries with lengths outside the 550 bp to 1000 bp range specified in the claims.
• —Does not cover workflows where the probability of homologous chromosome overlap exceeds 1%.

The clever bit

The innovation lies in using a high-density 5184-well plate to physically isolate DNA fragments before amplification, which statistically separates homologous chromosomes and simplifies the complex task of reassembling the genome later.

Why it matters

This method is significant for single-cell genomics and rare-sample analysis, where researchers have very little genetic material to work with. By minimizing the overlap of paternal and maternal chromosomes in a single well, it reduces errors in phasing, which is the process of determining which genetic variants belong to which chromosome. This is critical for clinical diagnostics and research into complex genetic diseases.

Real-world examples

• 1.Single-cell genomic sequencing workflows
• 2.Rare clinical sample genetic analysis
• 3.High-throughput DNA library preparation systems