Evidence map›Paper›PMID 29579126›Full record

ArticlePloS one2018

Improved methods of DNA extraction from human spermatozoa that mitigate experimentally-induced oxidative DNA damage.

Miguel J Xavier, Brett Nixon, Shaun D Roman, Robert John Aitken

Open access · goldAbstract read
In one paragraph

Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
1.0field-weighted citation impact, top 22% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

8 citing papers in PubMed, 13 citations in OpenAlex.

  1. Observational
  2. Article
  3. Article
  4. Review
  5. Article
  6. Impact of Heat Stress on Bovine Sperm Quality and Competence.Animals : an open access journal from MDPI · 2022
    Review
  7. Article
  8. Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

4 authors at 1 institution in 1 country.

Miguel J XavierReproductive Science Group, Faculty of Science, The University of Newcastle, Callaghan, NSW, Australia.ORCID 0000-0003-0709-7223
Brett NixonReproductive Science Group, Faculty of Science, The University of Newcastle, Callaghan, NSW, Australia.
Shaun D RomanReproductive Science Group, Faculty of Science, The University of Newcastle, Callaghan, NSW, Australia.
Robert John AitkenReproductive Science Group, Faculty of Science, The University of Newcastle, Callaghan, NSW, Australia.
University of Newcastle Australia · AU

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

Current approaches for DNA extraction and fragmentation from mammalian spermatozoa provide several challenges for the investigation of the oxidative stress burden carried in the genome of male gametes. Indeed, the potential introduction of oxidative DNA damage induced by reactive oxygen species, reducing agents (dithiothreitol or beta-mercaptoethanol), and DNA shearing techniques used in the preparation of samples for chromatin immunoprecipitation and next-generation sequencing serve to cofound the reliability and accuracy of the results obtained. Here we report optimised methodology that minimises, or completely eliminates, exposure to DNA damaging compounds during extraction and fragmentation procedures. Specifically, we show that Micrococcal nuclease (MNase) digestion prior to cellular lysis generates a greater DNA yield with minimal collateral oxidation while randomly fragmenting the entire paternal genome. This modified methodology represents a significant improvement over traditional fragmentation achieved via sonication in the preparation of genomic DNA from human spermatozoa for downstream applications, such as next-generation sequencing. We also present a redesigned bioinformatic pipeline framework adjusted to correctly analyse this form of data and detect statistically relevant targets of oxidation.

Indexed as

DNA DamageAdultDithiothreitolDNADNA FragmentationGenome, HumanHumansImmunoprecipitationLiquid-Liquid ExtractionMaleMercaptoethanolMicrococcal NucleaseMiddle AgedOxidative StressReactive Oxygen SpeciesShear StrengthDithiothreitolDNAMercaptoethanolMicrococcal NucleaseReactive Oxygen Species

Identifiers

PMID29579126
PMCPMC5868848
OpenAlexW2790516916

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.