Construction of high coverage whole-genome sequencing libraries from single colon crypts without DNA extraction or whole-genome amplification.

ABSTRACT:

Objective

Comprehensive and reliable genome-wide variant analysis of a small number of cells has been challenging due to genome coverage bias, PCR over-cycling, and the requirement of expensive technologies. To comprehensively identify genome alterations in single colon crypts that reflect genome heterogeneity of stem cells, we developed a method to construct whole-genome sequencing libraries from single colon crypts without DNA extraction, whole-genome amplification, or increased PCR enrichment cycles.

Results

We present post-alignment statistics of 81 single-crypts (each contains four- to eight-fold less DNA than the requirement of conventional methods) and 16 bulk-tissue libraries to demonstrate the consistent success in obtaining reliable coverage, both in depth (≥ 30X)

SUBMITTER: Manojlovic Z 

PROVIDER: S-EPMC10142246 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature