Popadin, K. Y. | bioRxiv Science

bioRxiv2024

Harnessing Stress: balancing the Burden of Slightly-Deleterious Variants by a Handicap

Numerous empirical studies have revealed epistatic interactions among deleterious variants. In this paper, we assume such interactions are widespread and analyze the resulting shift in mutational burden and average population fitness following the introduction of a strong and universal stress (hereafter "handicap"). We demonstrate that organisms with a low burden of slightly-deleterious variants (SDVs) are more likely to survive exposure to a handicap, whether genetic or environmental, leading to a purifying effect on the population. We further discuss the potential applications of harnessing such interactions for evolutionary and population studies as well as for population management.

genetics↗

bioRxiv2022

Isolation of highly purified genomic material from mitochondria of muscle tissue cells

In this work, we adapted a method for isolation of a highly purified fraction of mitochondrial DNA from muscle tissues suitable for further sample preparation of libraries without an amplification step for sequencing tasks on various NGS platforms and a method for evaluating the purity of DNA from contamination by nuclear genome regions. We optimized several techniques7,8 for enrichment of the mitochondrial fractions and purifying mtDNA. Here, we describe a protocol that allows getting from 80-100 mg of muscle tissues up to 1000 ng mtDNA, almost free from impurities of RNA and fragments of the nuclear genome.To assess the degree of purity of human mtDNA fraction from impurities of the nuclear genome, we adapted the PCR-screening technique7 for the beta-actin gene region and AluSx-repeats in the human genome. This methodology avoids false-heteroplasmy calls (PCR biases or NUMT contamination) that occur when long-range PCR amplification is used for mtDNA enrichment.

molecular biology↗