RT Journal Article SR Electronic T1 “Midi-metagenomics”: A novel approach for cultivation independent microbial genome reconstruction from environmental samples JF bioRxiv FD Cold Spring Harbor Laboratory SP 2023.01.26.525644 DO 10.1101/2023.01.26.525644 A1 John Vollmers A1 Maximiano Correa Cassal A1 Anne-Kristin Kaster YR 2023 UL http://biorxiv.org/content/early/2023/01/26/2023.01.26.525644.abstract AB As the majority of environmental microbial organisms still evade cultivation attempts, genomic insights into many taxa are limited to cultivation-independent approaches. However, current methods of metagenomic binning and single cell genome sequencing have individual drawbacks, which can limit the quality as well as completeness of the reconstructed genomes. Current attempts to combine both approaches still use whole genome amplification techniques which are known to be prone to bias. Here we propose a novel approach for the purpose of metagenomic genome reconstructions, that utilizes the potential of fluorescence-activated cell sorting (FACS) for targeted enrichment and depletion of different cell types to create distinct cell fractions of sufficient size circumvent amplification. By distributing sequencing efforts over these fractions as well as the original sample, co-assemblies become highly optimized for co-abundance variation based binning approaches. “Midi-metagenomics” enables accurate metagenome assembled genome (MAG) reconstruction from individual sorted samples with higher quality than co-assembly of multiple distinct samples and has potential for the targeted enrichment and sequencing of uncultivated organism of interest.Competing Interest StatementThe authors have declared no competing interest.