Literature DB >> 20041853

Nucleic acid contamination of glycogen used in nucleic acid precipitation and assessment of linear polyacrylamide as an alternative co-precipitant.

Andrea Bartram1, Calvin Poon, Josh Neufeld.   

Abstract

Molecular-grade glycogen is widely used to recover nanogram or picogram quantities of DNA and RNA across molecular biology applications in the life sciences. As a result, its purity is critical to obtain reliable results. Using agarose gel electrophoresis, we detected pg/microL (DNA) to ng/microL (RNA) concentrations of nucleic acid in two of the nine glycogen samples obtained from commercial suppliers. Denaturing gradient gel electrophoresis of 16S rRNA gene PCR-amplified products indicated that an additional two samples contained detectable contamination. We also tested a synthetic polymer co-precipitant, linear polyacrylamide (LPA); none of the four samples tested with LPA were detectably contaminated. The partial 16S rRNA gene sequence associated with the contaminated samples of the shellfish-derived glycogen was nearly identical to the sequence of Actinobacteria lwoffii, which has been isolated from mussels previously. By testing the recovery of low-nanogram amounts of DNA with multiple precipitants and simulated experimental conditions, we demonstrated that LPA was a preferable co-precipitant for sensitive protocols.

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Year:  2009        PMID: 20041853     DOI: 10.2144/000113276

Source DB:  PubMed          Journal:  Biotechniques        ISSN: 0736-6205            Impact factor:   1.993


  14 in total

1.  DNA stable-isotope probing (DNA-SIP).

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2.  Butyric acid- and dimethyl disulfide-assimilating microorganisms in a biofilter treating air emissions from a livestock facility.

Authors:  Anja Kristiansen; Sabine Lindholst; Anders Feilberg; Per H Nielsen; Josh D Neufeld; Jeppe L Nielsen
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3.  Genetic structure of the early Hungarian conquerors inferred from mtDNA haplotypes and Y-chromosome haplogroups in a small cemetery.

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Journal:  Mol Genet Genomics       Date:  2016-11-01       Impact factor: 3.291

4.  A method for extracting and characterizing RNA from urine: For downstream PCR and RNAseq analysis.

Authors:  Kun Zhou; Monique A Spillman; Kian Behbakht; Julia M Komatsu; Juan E Abrahante; Douglas Hicks; Brent Schotl; Evan Odean; Kenneth L Jones; Michael W Graner; Lynne T Bemis
Journal:  Anal Biochem       Date:  2017-08-10       Impact factor: 3.365

5.  Recovery comparison of two virus concentration methods from wastewater using cell culture and real-time PCR.

Authors:  Hasna Amdiouni; Leena Maunula; Kawtar Hajjami; Abdellah Faouzi; Abdelaziz Soukri; Jalal Nourlil
Journal:  Curr Microbiol       Date:  2012-07-06       Impact factor: 2.188

6.  Got Glycogen?: Development and Multispecies Validation of the Novel Preserve, Precipitate, Lyse, Precipitate, Purify (PPLPP) Workflow for Environmental DNA Extraction from Longmire's Preserved Water Samples.

Authors:  Richard C Edmunds; Damien Burrows
Journal:  J Biomol Tech       Date:  2020-12

7.  Aquarium nitrification revisited: Thaumarchaeota are the dominant ammonia oxidizers in freshwater aquarium biofilters.

Authors:  Laura A Sauder; Katja Engel; Jennifer C Stearns; Andre P Masella; Richard Pawliszyn; Josh D Neufeld
Journal:  PLoS One       Date:  2011-08-16       Impact factor: 3.240

8.  Expression and rhythmic modulation of circulating microRNAs targeting the clock gene Bmal1 in mice.

Authors:  Vikram R Shende; Marianna M Goldrick; Suchitra Ramani; David J Earnest
Journal:  PLoS One       Date:  2011-07-22       Impact factor: 3.240

9.  Methodological considerations for measuring biofluid-based microRNA biomarkers.

Authors:  Brian N Chorley; Elnaz Atabakhsh; Graeme Doran; Jean-Charles Gautier; Heidrun Ellinger-Ziegelbauer; David Jackson; Tatiana Sharapova; Peter S T Yuen; Rachel J Church; Philippe Couttet; Roland Froetschl; James McDuffie; Victor Martinez; Parimal Pande; Lauren Peel; Conor Rafferty; Frank J Simutis; Alison H Harrill
Journal:  Crit Rev Toxicol       Date:  2021-05-26       Impact factor: 6.184

10.  Comparison of Methods for miRNA Extraction from Plasma and Quantitative Recovery of RNA from Cerebrospinal Fluid.

Authors:  Melissa A McAlexander; Maggie J Phillips; Kenneth W Witwer
Journal:  Front Genet       Date:  2013-05-16       Impact factor: 4.599

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