Extraction of nucleic acids from bacterial spores using bead‐based mechanical lysis on a plastic chip

Geißler, Matthias; Beauregard, Julie A.; Charlebois, Isabelle; Isabel, Sandra; Normandin, François; Voisin, B.; Boissinot, Maurice; Bergeron, Michel G.; Veres, Teodor

doi:10.1002/elsc.201000132

Cited by 10 publications

(6 citation statements)

References 21 publications

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“…While the statement of ratio of sample to buffer at the ratio of 1:5 was found to be optimal in protein extraction of monoclonal antibodies (Gottschalk, 2014), this result seems contradictive with the result of this experiment. Geissler et al (2011) agree that the ratio of 5 mL buffer per gram can be the starting point to avoid the loss of protein activity or nonspecific binding to containers. However, then again, the ratio of buffer has to be lower down, such as 1:1 or 1:2, if concentrated sample is essential.…”

Section: Discussionmentioning

confidence: 91%

Preliminary Study of Buffer Ratio in Protein Extraction from Placental Cotyledons of Kedah-Kelantan Cattle

Tee

Mat

Adam

et al. 2020

Trop. Anim. Sci. J.

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Protein extraction is a preliminary step of protein purification which mainly focus on maximization of total protein yield. The heterogeneous properties cause diversification of protein; therefore, there is no absolute protocol in protein extraction. The ratio of buffer gives different protein concentrations in different types of mammalian tissues, and this condition leads to the study of optimization of buffer ratio to obtain a better total protein yield. The objectives of this study were to compare the total protein yield based on three different ratios of buffer used. The phosphate buffer saline (PBS), radioimmunoprecipitation assay (RIPA) buffer, and RIPA buffer with the addition of protease inhibitor (Pi) were used with the ratios of 1:1, 1:3, and 1:5. Fetal cotyledons removed from the placenta have undergone mechanical disruption, incubation, sonication, and centrifugation. The supernatant was retained and quantified with Bradford assay to determine the total protein yield based on the standard curve of bovine serum albumin (BSA). There was a statistically significant different between buffer ratio (p<0.5) in RIPA and RIPA with addition of protease inhibitor buffers. RIPA buffer with the ration of 1:1 gave the best total protein yield (194.880±15.089 mg/g). As a conclusion, there was a significant interaction between buffer types and have greatly enhanced the total protein yield obtained from placental cotyledons of Kedah-Kelantan cattle.

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Section: Discussionmentioning

confidence: 91%

Preliminary Study of Buffer Ratio in Protein Extraction from Placental Cotyledons of Kedah-Kelantan Cattle

Tee

Mat

Adam

et al. 2020

Trop. Anim. Sci. J.

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“…B. atrophaeus subsp. globigii CCRI-9827 spores (at a 0.5 McFarland standard) were labeled with 0.02% fluorescein isothiocyanate (FITC) as described previously (3). The penetration and release of fluorescent B. atrophaeus spores in 5-m-and 0.45-m-poresize filters were evaluated by using an Olympus FV300 confocal laser scanning microscope (Olympus Canada, Markham, Ontario, Canada) with an argon-ion laser for excitation at 488 nm.…”

Section: Methodsmentioning

confidence: 99%

Rapid Filtration Separation-Based Sample Preparation Method for Bacillus Spores in Powdery and Environmental Matrices

Isabel

Boissinot

Charlebois

et al. 2012

Appl Environ Microbiol

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Authorities frequently need to analyze suspicious powders and other samples for biothreat agents in order to assess environmental safety. Numerous nucleic acid detection technologies have been developed to detect and identify biowarfare agents in a timely fashion. The extraction of microbial nucleic acids from a wide variety of powdery and environmental samples to obtain a quality level adequate for these technologies still remains a technical challenge. We aimed to develop a rapid and versatile method of separating bacteria from these samples and then extracting their microbial DNA. Bacillus atrophaeus subsp. globigii was used as a simulant of Bacillus anthracis. We studied the effects of a broad variety of powdery and environmental samples on PCR detection and the steps required to alleviate their interference. With a benchmark DNA extraction procedure, 17 of the 23 samples investigated interfered with bacterial lysis and/or PCR-based detection. Therefore, we developed the dual-filter method for applied recovery of microbial particles from environmental and powdery samples (DARE). The DARE procedure allows the separation of bacteria from contaminating matrices that interfere with PCR detection. This procedure required only 2 min, while the DNA extraction process lasted 7 min, for a total of <10 min. This sample preparation procedure allowed the recovery of cleaned bacterial spores and relieved detection interference caused by a wide variety of samples. Our procedure was easily completed in a laboratory facility and is amenable to field application and automation.

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“…For continuous-flow devices, glass or plastic is more used, since a low thermal conductivity is desirable for the realization of different temperature zones in the chip [ 67 ]. Therefore, many microfluidic devices for biological fluid analysis are made from PDMS or PMMA [ 18 , 19 , 39 , 88 , 163 , 164 ], although glass or silicon is also utilized [ 31 , 58 , 65 , 152 , 165 ].…”

Section: Chip Materials For Dna Analysismentioning

confidence: 99%

Microfluidic Devices for Forensic DNA Analysis: A Review

et al. 2016

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Microfluidic devices may offer various advantages for forensic DNA analysis, such as reduced risk of contamination, shorter analysis time and direct application at the crime scene. Microfluidic chip technology has already proven to be functional and effective within medical applications, such as for point-of-care use. In the forensic field, one may expect microfluidic technology to become particularly relevant for the analysis of biological traces containing human DNA. This would require a number of consecutive steps, including sample work up, DNA amplification and detection, as well as secure storage of the sample. This article provides an extensive overview of microfluidic devices for cell lysis, DNA extraction and purification, DNA amplification and detection and analysis techniques for DNA. Topics to be discussed are polymerase chain reaction (PCR) on-chip, digital PCR (dPCR), isothermal amplification on-chip, chip materials, integrated devices and commercially available techniques. A critical overview of the opportunities and challenges of the use of chips is discussed, and developments made in forensic DNA analysis over the past 10–20 years with microfluidic systems are described. Areas in which further research is needed are indicated in a future outlook.

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Extraction of nucleic acids from bacterial spores using bead‐based mechanical lysis on a plastic chip

Cited by 10 publications

References 21 publications

Preliminary Study of Buffer Ratio in Protein Extraction from Placental Cotyledons of Kedah-Kelantan Cattle

Preliminary Study of Buffer Ratio in Protein Extraction from Placental Cotyledons of Kedah-Kelantan Cattle

Rapid Filtration Separation-Based Sample Preparation Method for Bacillus Spores in Powdery and Environmental Matrices

Microfluidic Devices for Forensic DNA Analysis: A Review

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