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Polytene chromosomes are giant interphase chromosomes with several DNA strands placed side by side. They were discovered in the year 1881 by Balbiani in salivary glands, intestine, muscles, malpighian tubules, and hypoderm of larvae Chironomus plumosus. Hence, these are also called "Salivary gland chromosomes." These are found in insects of the order Diptera and Collembola; in certain organs of mammals; and synergids, antipodes of flowering plants. Polytene chromosomes are also regularly...
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Manipulation of Ploidy in Caenorhabditis elegans
07:54

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Published on: March 15, 2018

Artificial polyploidy improves bacterial single cell genome recovery.

Armand E K Dichosa1, Michael S Fitzsimons, Chien-Chi Lo

  • 1Department of Energy Joint Genome Institute, Los Alamos National Laboratory, Los Alamos, New Mexico, United States of America.

Plos One
|June 6, 2012
PubMed
Summary

Researchers developed a novel method to improve single-cell genomics (SCG) by inducing polyploidy in bacteria. This technique enhances genomic recovery and reduces amplification bias for complete genome sequencing from single cells.

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Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Single-cell genomics (SCG) aims to sequence entire genomes from individual cells.
  • Previous SCG studies faced challenges like DNA fragmentation and amplification bias, limiting complete genome acquisition, especially in bacteria.
  • Organisms with smaller genomes, such as bacteria and archaea, have been the primary focus for SCG.

Purpose of the Study:

  • To investigate an artificial method for inducing polyploidy in Bacillus subtilis ATCC 6633.
  • To improve the performance of genomic sequencing from a single bacterial cell.
  • To overcome limitations of DNA breakages and amplification bias in SCG.

Main Methods:

  • Inhibited the bacterial cytoskeleton protein FtsZ in B. subtilis using PC190723.
  • Induced polyploidy, resulting in larger, undivided single cells with multiple genome copies.
  • Utilized quantitative PCR (qPCR) assays on sorted cells to assess DNA content and amplification bias.

Main Results:

  • Successfully generated larger, undivided B. subtilis cells with multiple genomes.
  • Sorted cells exhibited higher DNA content compared to untreated cells.
  • Demonstrated reduced amplification bias and greater genomic recovery in polyploid cells.

Conclusions:

  • The developed method shows potential for obtaining nearly complete bacterial genomes from single cells.
  • This approach offers significant promise for exploring the genomic novelty of millions of uncultured bacterial species.
  • Artificial polyploidy, combined with cell sorting, may enable the recovery of specific organisms and their genomes.