Extraction and Amplification of Genomic DNA from Human Blood on Nanoporous Aluminum Oxide Membranes

Elgort, Marc G.; Herrmann, Mark G.; Erali, Maria; Durtschi, Jacob D.; Voelkerding, Karl V.; Smith, R.E.

doi:10.1373/clinchem.2004.035162

Cited by 22 publications

(19 citation statements)

References 8 publications

(2 reference statements)

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“…Membranes play an important role in science and technology, such as the applications of ion exchange membranes in separation [ 1 , 2 , 3 ], wastewater treatment [ 2 , 3 , 4 ], food, and bio-technology [ 3 , 4 ]; nanoporous membranes in blood purification [ 5 ], separation and gas removal [ 6 , 7 ], virus filtration [ 8 ] and DNA/gene detection [ 9 , 10 ]; liquid crystal polymer membranes in optics, electronics, display devices, and bio-technology [ 11 , 12 , 13 , 14 ]; honeycomb films in anti-bacterial activity [ 15 , 16 ], cell/bacteria culture [ 17 , 18 ], protein adsorption [ 19 ], tissue engineering [ 20 ], lab-on-fiber technology [ 21 ], electrowetting display [ 22 ], and water/oil separation [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

Breath Figure Method for Construction of Honeycomb Films

et al. 2015

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Honeycomb films with various building units, showing potential applications in biological, medical, physicochemical, photoelectric, and many other areas, could be prepared by the breath figure method. The ordered hexagonal structures formed by the breath figure process are related to the building units, solvents, substrates, temperature, humidity, air flow, and other factors. Therefore, by adjusting these factors, the honeycomb structures could be tuned properly. In this review, we summarized the development of the breath figure method of fabricating honeycomb films and the factors of adjusting honeycomb structures. The organic-inorganic hybrid was taken as the example building unit to discuss the preparation, mechanism, properties, and applications of the honeycomb films.

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Section: Introductionmentioning

confidence: 99%

Breath Figure Method for Construction of Honeycomb Films

et al. 2015

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“…Therefore, the efficiency of a DNA extraction protocol will be affected by its robustness and ability to yield bulk amount of clean and unblemished genomic DNA. A number of DNA extraction protocols have already been published by several workers around the globe, which were verified to be reproducible and efficient in yielding sufficient amount of high quality DNA . However, use of expensive enzymes and toxic solvents in these extraction procedures raised questions on their competence.…”

Section: Discussionmentioning

confidence: 99%

“…Many protocols have been published regarding DNA isolation from blood . Some of these published protocols applied enzymes and organic solvents for yielding high quality DNA, devoid of PCR inhibitors, while others including salting out procedure targeted toward higher DNA yields .…”

Section: Introductionmentioning

confidence: 99%

A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples

Guha

Das

Dutta

et al. 2017

Clinical Laboratory Analysis

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Background Different methods available for extraction of human genomic DNA suffer from one or more drawbacks including low yield, compromised quality, cost, time consumption, use of toxic organic solvents, and many more. Herein, we aimed to develop a method to extract DNA from 500 μL of fresh or frozen human blood. Methods Five hundred microliters of fresh and frozen human blood samples were used for standardization of the extraction procedure. Absorbance at 260 and 280 nm, respectively, (A260/A280) were estimated to check the quality and quantity of the extracted DNA sample. Qualitative assessment of the extracted DNA was checked by Polymerase Chain reaction and double digestion of the DNA sample. Results Our protocol resulted in average yield of 22±2.97 μg and 20.5±3.97 μg from 500 μL of fresh and frozen blood, respectively, which were comparable to many reference protocols and kits. Conclusion Besides yielding bulk amount of DNA, our protocol is rapid, economical, and avoids toxic organic solvents such as Phenol. Due to unaffected quality, the DNA is suitable for downstream applications. The protocol may also be useful for pursuing basic molecular researches in laboratories having limited funds.

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“…17 AOMs can also inhibit PCR to some degree, but the basic pH of the master mix along with adding BSA and extra Taq polymerase to the reaction mixture will release nucleic acids bound to the membrane and minimize the inhibition. 16, 17, 23, 26 …”

Section: Introductionmentioning

confidence: 99%

A microfluidic chip integrating DNA extraction and real-time PCR for the detection of bacteria in saliva

et al. 2013

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A microfluidic chip integrating DNA extraction, amplification, and detection for the identification of bacteria in saliva is described. The chip design integrated a monolithic aluminum oxide membrane (AOM) for DNA extraction with seven parallel reaction wells for real-time polymerase chain reaction (rtPCR) amplification of the extracted DNA. Samples were first heated to lyse target organisms and then added to the chip and filtered through the nanoporous AOM to extract the DNA. PCR reagents were added to each of the wells and the chip was thermocycled. Identification of Streptococcus mutans in a saliva sample is demonstrated along with the detection of 300 fg (100–125 copies) of both methicillin-susceptible Staphylococcus aureus (MSSA) and methicillin-resistant S. aureus (MRSA) genomic DNA (gDNA) spiked into a saliva sample. Multiple target species and strains of bacteria can be simultaneously identified in the same sample by varying the primers and probes used in each of the seven reaction wells. In initial tests, as little as 30 fg (8–12 copies) of MSSA gDNA in buffer has been successfully amplified and detected with this device.

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Extraction and Amplification of Genomic DNA from Human Blood on Nanoporous Aluminum Oxide Membranes

Cited by 22 publications

References 8 publications

Breath Figure Method for Construction of Honeycomb Films

Breath Figure Method for Construction of Honeycomb Films

A rapid and efficient DNA extraction protocol from fresh and frozen human blood samples

A microfluidic chip integrating DNA extraction and real-time PCR for the detection of bacteria in saliva

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