Assessing recovery rates of distinct exogenous controls for gDNA extraction efficiency using phenol-chloroform or silica-column based extractions.

J Microbiol Methods

Laboratório of Research in Biofilms Rosário Oliveira (LIBRO), Centre of Biological Engineering (CEB), University of Minho, Campus de Gualtar, 4710-057 Braga, Portugal; LABBELS -Associate Laboratory, Braga & Guimarães, Portugal. Electronic address:

Published: December 2022


Category Ranking

98%

Total Visits

921

Avg Visit Duration

2 minutes

Citations

20

Article Abstract

Assessment of genomic DNA (gDNA) extraction efficiency is required for accurate bacterial quantification by qPCR. Exogenous DNA molecules are often added after bacterial cultures are lysed, but before DNA purification steps, to determine extraction efficiency. Herein we found that different exogenous DNA controls have different recovery rates, suggesting distinct DNA extraction efficiencies. Recovery rates are also affected by the gDNA extraction method being more affected in silica-based columns than in phenol-chloroform extraction. Overall, we determined that the use of long DNA fragments, such as gDNA, as exogenous controls have a higher recovery rate than use of smaller size DNA molecules.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10804838PMC
http://dx.doi.org/10.1016/j.mimet.2022.106607DOI Listing

Publication Analysis

Top Keywords

recovery rates
12
gdna extraction
12
extraction efficiency
12
exogenous controls
8
exogenous dna
8
dna molecules
8
dna
7
extraction
6
assessing recovery
4
rates distinct
4

Similar Publications

Background: People with opioid use disorder (OUD) often exhibit high rates of nonprescribed drug use and low retention on buprenorphine. This study tested the feasibility, acceptability, and preliminary efficacy of an intervention combining peer recovery coaching and CBT4CBT-buprenorphine (CBT4CBT+RC) to reduce nonprescribed drug use and increase buprenorphine retention.

Methods: A randomized trial conducted from December 15, 2020, to November 24, 2021, compared an 8-week CBT4CBT+RC intervention to treatment as usual (TAU).

View Article and Find Full Text PDF

To evaluate long‑term outcomes of corneal patch grafting (CPG) and to determine prognostic factors for anatomical and functional success. This retrospective study included 35 eyes from 35 patients who underwent CPG between April 2016 and September 2022 at Adana City Training and Research Hospital. Collected data included age, sex, preoperative and postoperative best-corrected visual acuity (BCVA), graft localization and size, anterior segment findings, graft survival, secondary surgical procedures, and rates of anatomical and functional success.

View Article and Find Full Text PDF

ObjectiveThe sedation protocol for flexible fiberoptic bronchoscopy has long been a matter of inconclusiveness. The aim of this study was to evaluate the safety and efficacy of remimazolam combined with alfentanil in flexible fiberoptic bronchoscopy and provide insights for optimizing clinical anesthesia strategies.MethodsThis study was a randomized, single-blind controlled trial.

View Article and Find Full Text PDF

In this study, a one-pot hydrothermal synthesis method was used to synthesize a novel gold-yttrium trimesic acid metal-organic framework (Au-Y-TMA MOF), demonstrating significant improvements over conventional single-metal MOFs, that is, yttrium trimesic acid (Y-TMA), in both supercapacitor applications and electrochemical antibiotic detection. The X-ray diffraction patterns of Au-Y-TMA confirmed the presence and impact of Au in the Y-TMA matrix, while field emission scanning electron microscopy (FE-SEM) images revealed a heterogeneous combination of gold nanoparticles (AuNPs) and Y-TMA, suggesting a nonuniform distribution and possible interaction. The developed half-cell supercapacitor exhibited a remarkable capacitance value of 1836 F/g at a current density of 5 A/g by galvanostatic charging-discharging (GCD) measurement.

View Article and Find Full Text PDF

Background: Achilles tendon ruptures are common, particularly in active individuals, and significantly affect function. Controversy persists over whether conservative or surgical treatment offers superior outcomes.

Objective: To compare conservative and surgical treatments for Achilles tendon rupture regarding rerupture rates, functional recovery, and complication incidence.

View Article and Find Full Text PDF