A Single-Tube Nucleic Acid Extraction, Amplification, and Detection Method Using Aluminum Oxide

Dames, Shale; Bromley, L. Kathryn; Herrmann, Mark G.; Elgort, Marc G.; Erali, Maria; Smith, R.E.; Voelkerding, Karl V.

doi:10.2353/jmoldx.2006.040398

Cited by 16 publications

(14 citation statements)

References 12 publications

(13 reference statements)

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“…However, the binding of nucleic acids to AAO in the presence of guanidinium appears to be irreversible (21). This phenomenon might be exploited for solid-phase amplifications (20), but for our application it is a possible source of loss.…”

Section: Discussionmentioning

confidence: 99%

“…We chose the salting-out extraction procedure, rather than one of the popular guanidinium-based extraction methods, because guanidinium salts have been reported to cause irreversible binding of DNA to AAO (20,21). We later described a modification of our filter-extraction protocol (22) in which extraction buffer is back flushed through the membrane rather than being pushed through in the same direction as the sample.…”

mentioning

confidence: 99%

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Variables Influencing Extraction of Nucleic Acids from Microbial Plankton (Viruses, Bacteria, and Protists) Collected on Nanoporous Aluminum Oxide Filters

Mueller

Culley

Steward

2014

Appl Environ Microbiol

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Anodic aluminum oxide (AAO) filters have high porosity and can be manufactured with a pore size that is small enough to quantitatively capture viruses. These properties make the filters potentially useful for harvesting total microbial communities from water samples for molecular analyses, but their performance for nucleic acid extraction has not been systematically or quantitatively evaluated. In this study, we characterized the flux of water through commercially produced nanoporous (0.02 m) AAO filters (Anotop; Whatman) and used isolates (a virus, a bacterium, and a protist) and natural seawater samples to test variables that we expected would influence the efficiency with which nucleic acids are recovered from the filters. Extraction chemistry had a significant effect on DNA yield, and back flushing the filters during extraction was found to improve yields of high-molecularweight DNA. Using the back-flush protocol, the mass of DNA recovered from microorganisms collected on AAO filters was >100% of that extracted from pellets of cells and viruses and 94% ؎ 9% of that obtained by direct extraction of a liquid bacterial culture. The latter is a minimum estimate of the relative recovery of microbial DNA, since liquid cultures include dissolved nucleic acids that are retained inefficiently by the filter. In conclusion, we demonstrate that nucleic acids can be extracted from microorganisms on AAO filters with an efficiency similar to that achievable by direct extraction of microbes in suspension or in pellets. These filters are therefore a convenient means by which to harvest total microbial communities from multiple aqueous samples in parallel for subsequent molecular analyses.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Variables Influencing Extraction of Nucleic Acids from Microbial Plankton (Viruses, Bacteria, and Protists) Collected on Nanoporous Aluminum Oxide Filters

Mueller

Culley

Steward

2014

Appl Environ Microbiol

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“…Диагноз генитального герпеса ставился на основании анамнеза заболевания, типичных высыпаний, выявления антигена ВПГ 2 типа в ИФА, в тест-системе «Герпескрин» (Ниармедик плюс, Россия), а также с помощью ПЦР (in house), с использованием праймеров, специфичных для Herpes simplex, тип 2 -5`-GTACAGACCTTCGGAGG-3` и 5`-CGCTTCATCATGGGC-3` [11]. Использовали следующий режим амплификации (40 циклов): 94°C -30 с., 60°С -40 с., 72°С -50 с. на амплификаторе «Терцик» (Россия).…”

Section: Introductionunclassified

Современные Особенности Проявлений Генитального Герпеса, Хламидиоза И Трихомоноза

Nagorniy¹

2022

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“…3,4) This method is rapid and requires few manipulations. Single-tube real-time PCR methods, using an aluminum oxide filter and a heated guanidine solution, have also been attempted.…”

mentioning

confidence: 99%

“…Single-tube real-time PCR methods, using an aluminum oxide filter and a heated guanidine solution, have also been attempted. 3,4) However, these extraction processes required special handling, such as pressure filtration and dry evaporation of samples. In addition, these methods could extract total DNA from a sample but could not specifically isolate target DNA.…”

mentioning

confidence: 99%

Real-Time PCR Method Using Capturing Oligo-Immobilized PCR Tubes to Determine the Specific Gene for Soybean and Genetically Modified Soybean in Food Matrices

Harikai¹,

Saito²,

Abe³

et al. 2008

Bioscience, Biotechnology, and Biochemistry

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A Single-Tube Nucleic Acid Extraction, Amplification, and Detection Method Using Aluminum Oxide

Cited by 16 publications

References 12 publications

Variables Influencing Extraction of Nucleic Acids from Microbial Plankton (Viruses, Bacteria, and Protists) Collected on Nanoporous Aluminum Oxide Filters

Variables Influencing Extraction of Nucleic Acids from Microbial Plankton (Viruses, Bacteria, and Protists) Collected on Nanoporous Aluminum Oxide Filters

Современные Особенности Проявлений Генитального Герпеса, Хламидиоза И Трихомоноза

Real-Time PCR Method Using Capturing Oligo-Immobilized PCR Tubes to Determine the Specific Gene for Soybean and Genetically Modified Soybean in Food Matrices

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