A facile one-step method for cell lysis and DNA extraction of waterborne pathogens using a microchip

Kamat, Vivek; Pandey, Sulaxna; Paknikar, Kishore M.; Bodas, Dhananjay

doi:10.1016/j.bios.2017.07.040

Cited by 27 publications

(15 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As starting point, the approaches that were developed for cell lysis are available. They are based on either chemical [ 204 ] or physical (electrical [ 205 , 206 ], magnetic [ 207 ]) methods, and may be potentially applied also to the membrane of MVs or exosomes. Commonly used chemical lysis buffers include sodium dodecyl sulfate (SDS), proteinase K or Triton X-100 even though a differential efficacy has been reported on various target subpopulations: for example, MVs and apoptotic bodies were found to be more sensitive to detergent lysis than exosomes [ 2 ].…”

Section: Exploring the Content Of Vesiclesmentioning

confidence: 99%

“…When dealing with nucleic acids, which are also conveyed from EVs and may work as carrier for mutations with a role in carcinogenesis, a convenient strategy is typically to exploit amplification methods for generating multiple copies of a target nucleic acid, either through standard PCR cycles or alternative approaches, such as isothermal amplification [ 207 , 212 ] and subsequently analyse the sequence looking for specific genes or point mutations. However, off-chip nucleic acid-based detection relies on complex analytical procedures and requires a relatively long time with multiple steps, expensive instruments and reagents, and trained personnel.…”

Section: Exploring the Content Of Vesiclesmentioning

confidence: 99%

See 1 more Smart Citation

Lab-on-Chip for Exosomes and Microvesicles Detection and Characterization

Chiriaco

Bianco

Nigro

et al. 2018

Sensors

128

View full text Add to dashboard Cite

Interest in extracellular vesicles and in particular microvesicles and exosomes, which are constitutively produced by cells, is on the rise for their huge potential as biomarkers in a high number of disorders and pathologies as they are considered as carriers of information among cells, as well as being responsible for the spreading of diseases. Current methods of analysis of microvesicles and exosomes do not fulfill the requirements for their in-depth investigation and the complete exploitation of their diagnostic and prognostic value. Lab-on-chip methods have the potential and capabilities to bridge this gap and the technology is mature enough to provide all the necessary steps for a completely automated analysis of extracellular vesicles in body fluids. In this paper we provide an overview of the biological role of extracellular vesicles, standard biochemical methods of analysis and their limits, and a survey of lab-on-chip methods that are able to meet the needs of a deeper exploitation of these biological entities to drive their use in common clinical practice.

show abstract

Section: Exploring the Content Of Vesiclesmentioning

confidence: 99%

Section: Exploring the Content Of Vesiclesmentioning

confidence: 99%

Lab-on-Chip for Exosomes and Microvesicles Detection and Characterization

Chiriaco

Bianco

Nigro

et al. 2018

Sensors

128

View full text Add to dashboard Cite

show abstract

“…Although the addition of alkali can also dissolve L-tyrosine [ 3 ], the E. coli cells are lysed, after which a large number of cell fragments, nucleic acids, and proteins are released [ 11 ]. Following are few drawbacks of cell lysis: (1) The broth becomes very viscous; thick fermented broth, like the thick mucus of a flu patient, is very difficult to handle later.…”

Section: Resultsmentioning

confidence: 99%

A new method to recover L-tyrosine from E. coli fermentation broth

JingYang

Chen

et al. 2020

Bioengineered

View full text Add to dashboard Cite

Although the production of L-tyrosine by recombinant Escherichia coli has been widely reported, L-tyrosine recovery from the fermentation broth has been rarely reported. Methods to recover L-tyrosine from the broth after alkaline lysis of the bacterial cells have been described. However, the broth becomes viscous and dark following cell lysis, making further extraction and purification difficult. Here, a new method for L-tyrosine extraction and purification from the fermentation broth without the lysis of bacteria is reported. First, acids, rather than bases, were used to dissolve L-tyrosine in the broth without causing lysis of the bacterial cells. E. coli cells in the broth were then removed through centrifugation. Activated carbon was then used to decolorize the supernatant containing L-tyrosine. Finally, sodium hydroxide was added to the clarified L-tyrosine solution for isoelectrocrystallization. L-tyrosine was obtained after filtration and drying. The recovery yield of L-tyrosine was 92%, and the purity was >98.5%, indicating high efficiency of the new method of L-tyrosine recovery from fermented broth. Furthermore, the method provides a reference for the extraction of guanosine, inosine, hypoxanthine, and other biological fermentation products.

show abstract

“…Purification and isolation of nucleotide samples like deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) is an important step for subsequent characterization and application of various biochemical reaction [1][2][3]. The common methods of purification of DNA include electrophoresis [4], high salt precipitation [5], spin column [6], and magnetic beads [7], etc.…”

Section: Introductionmentioning

confidence: 99%

Continuous Microfluidic Purification of DNA Using Magnetophoresis

et al. 2020

View full text Add to dashboard Cite

Automatic microfluidic purification of nucleic acid is predictable to reduce the input of original samples and improve the throughput of library preparation for sequencing. Here, we propose a novel microfluidic system using an external NdFeB magnet to isolate DNA from the polymerase chain reaction (PCR) mixture. The DNA was purified and isolated when the DNA-carrying beads transported to the interface of multi-laminar flow under the influence of magnetic field. Prior to the DNA recovery experiments, COMSOL simulations were carried out to study the relationship between trajectory of beads and magnet positions as well as fluid velocities. Afterwards, the experiments to study the influence of varying velocities and input of samples on the DNA recovery were conducted. Compared to experimental results, the relative error of the final position of beads is less than 10%. The recovery efficiency decreases with increase of input or fluid velocity, and the maximum DNA recovery efficiency is 98.4% with input of l00 ng DNA at fluid velocity of 1.373 mm/s. The results show that simulations significantly reduce the time for parameter adjustment in experiments. In addition, this platform uses a basic two-layer chip to realize automatic DNA isolation without any other liquid switch value or magnet controller.

show abstract

A facile one-step method for cell lysis and DNA extraction of waterborne pathogens using a microchip

Cited by 27 publications

References 26 publications

Lab-on-Chip for Exosomes and Microvesicles Detection and Characterization

Lab-on-Chip for Exosomes and Microvesicles Detection and Characterization

A new method to recover L-tyrosine from E. coli fermentation broth

Continuous Microfluidic Purification of DNA Using Magnetophoresis

Contact Info

Product

Resources

About