Methods and kits for intracellular amplification and sequencing (fluorescently labeled intracellular genomics (fling))

Abstract

Rare genotypes in bacterial or eukaryotic populations are essential for understanding adaptation, ecology, and disease, but assembling the full genome of rare cell types remains challenging. Bulk sequencing of mixed populations obscures rare genotypes and haplotype information, while random single-cell isolation is labor-intensive, particularly for rare cell types. Described here is a novel method that enriches cells with a known genetic marker, enabling efficient sequencing of their partial or full genome. This approach integrates intracellular genomic DNA amplification, Fluorescent In Situ Amplification (FISA), Fluorescence-Activated Cell Sorting (FACS), and whole-genome sequencing. It allows robust amplification and sequencing of single genes and entire genomes from formaldehyde-fixed cells, preserving cellular integrity and minimizing false positives. This versatile method provides a powerful tool for studying rare genotypes and genetic heterogeneity across applications in evolution, ecology, and medicine.

Dependencies are inferred from the claim text.