Erratum to “DNA, RNA, and Protein Extraction: The Past and the Present”

Tan, Siun Chee; Yiap, Beow Chin

doi:10.1155/2013/628968

Cited by 22 publications

(28 citation statements)

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“…When working with whole blood samples, cell lysis and RNase denaturation are required to obtain intact viral RNA (Tan and Yiap 2009). It has been shown that 80%–95% of the total viral RNA can be rapidly isolated from 200 μL of whole blood using an off-chip commercial method (ZR Whole-Blood RNA MiniPrep™), thus our sample preparation protocol is compatible with the fingerprick blood test (Anderson et al 1999).…”

Section: Discussionmentioning

confidence: 99%

Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus

Cai

Park

et al. 2017

Biosensors and Bioelectronics

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An automated microfluidic sample preparation multiplexer (SPM) has been developed and evaluated for Ebola virus detection. Metered air bubbles controlled by microvalves are used to improve bead-solution mixing thereby enhancing the hybridization of the target Ebola virus RNA with capture probes bound to the beads. The method uses thermally stable 4-formyl benzamide functionalized (4FB) magnetic beads rather than streptavidin coated beads with a high density of capture probes to improve the target capture efficiency. Exploiting an on-chip concentration protocol in the SPM and the single molecule detection capability of the antiresonant reflecting optical waveguide (ARROW) biosensor chip, a detection limit of 0.021 pfu/mL for clinical samples is achieved without target amplification. This RNA target capture efficiency is two orders of magnitude higher than previous results using streptavidin beads and the limit of detection (LOD) improves 10X. The wide dynamic range of this technique covers the whole clinically applicable concentration range. In addition, the current sample preparation time is ~1 hour which is eight times faster than previous work. This multiplexed, miniaturized sample preparation microdevice establishes a key technology that intended to develop next generation point-of-care (POC) detection system.

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Section: Discussionmentioning

confidence: 99%

Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus

Cai

Park

et al. 2017

Biosensors and Bioelectronics

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“…To avoid the contaminating the peak at 230 nm, we decided to use a solubilisation buffer devoid of guanidine salts. Therefore, we tested a procedure based on the use of SDS-P/C/I for an efficient cell lysis and RNase inactivation [15], coupled to TRIzol (Thermofisher Scientific) [8]. We used this procedure together with the subsequent RNA clean-up detailed in the RNeasy mini kit protocol (Qiagen).…”

Section: Comparison Of the Rna Extraction Methodsmentioning

confidence: 99%

Extraction of High Quality RNA from Cannabis sativa Bast Fibres: A Vademecum for Molecular Biologists

Guerriero

Mangeot-Peter

Hausman

et al. 2016

Fibers

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Abstract:In plants there is no universal protocol for RNA extraction, since optimizations are required depending on the species, tissues and developmental stages. Some plants/tissues are rich in secondary metabolites or synthesize thick cell walls, which hinder an efficient RNA extraction. One such example is bast fibres, long extraxylary cells characterized by a thick cellulosic cell wall. Given the economic importance of bast fibres, which are used in the textile sector, as well as in biocomposites as green substitutes of glass fibres, it is desirable to better understand their development from a molecular point of view. This knowledge favours the development of biotechnological strategies aimed at improving specific properties of bast fibres. To be able to perform high-throughput analyses, such as, for instance, transcriptomics of bast fibres, RNA extraction is a crucial and limiting step. We here detail a protocol enabling the rapid extraction of high quality RNA from the bast fibres of textile hemp, Cannabis sativa L., a multi-purpose fibre crop standing in the spotlight of research.

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“…In general, nucleic acid extraction procedures involve four fundamental steps: (a) lysis of cells or tissues; (b) denaturation of nucleoprotein complexes; (c) inactivation of nucleases (DNase in case of isolation of DNA and RNase in the case of RNA purification); and (d) removal of contaminants . More specifically, these extractive methods can isolate nucleic acids from other cellular biomolecules (such as proteins, lipids, and carbohydrates) and molecules that potentially interfere with enzymes employed in downstream processes, most of the times, polymerase chain reaction, reverse transcription‐polymerase chain reaction (i.e., DNA polymerase or RNA dependent DNA polymerase), or southern blots (i.e., antibody binding) . Usually, a quality control of nucleic acid extracted solutions is performed by means of UV absorption spectrophotometry.…”

Section: Introductionmentioning

confidence: 99%

“…For DNA, we used QIAamp DNA Mini Kit (Qiagen). It is a silica‐membrane‐based DNA purification method and exemplificative of a typical solid‐phase nucleic acid extraction, commonly followed in most of the commercial extraction kits . Instead, for RNA, we used the TRIzol RNA extraction protocol (Thermo Scientific), by using Isol‐RNA lysis reagent (5‐Prime).…”

Section: Introductionmentioning

confidence: 99%

The quality is in the eye of the beholder: The perspective of FTIR and UV resonant Raman spectroscopies on extracted nucleic acids

Zucchiatti

Latella

Birarda

et al. 2018

J Raman Spectroscopy

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The quality of a sample has always to be considered with respect to its purposes and to the technique used for assessing it. The aim of this work is to point out the most common interfering agents that vibrational spectroscopists may find when analyzing nucleic acids extracted from cellular cultures. Fourier transformed infrared and ultraviolet‐resonant Raman measurements have been carried out on DNA and RNA samples extracted from B16 cell cultures by standard protocols used in molecular biology. The routinely adopted quality control procedure, based on ultraviolet spectrophotometry absorption measurements, could only indicate the absence of protein, phenol, or other contaminants that absorb strongly at or near 280 nm. However, DNA and RNA samples of good quality from a biological perspective clearly showed the presence of chemicals interfering with the vibrational measurements, mainly ethanol and guanidinium salts. Here, we propose fast and inexpensive strategies for tailoring the extraction protocols in light of the requirements of both vibrational spectroscopies that allowed obtaining nucleic acid samples with ethanol concentration below the detection limit of both techniques and with a significantly reduced spectral interference from guanidinium salts.

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Erratum to “DNA, RNA, and Protein Extraction: The Past and the Present”

Cited by 22 publications

References 0 publications

Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus

Multiplexed efficient on-chip sample preparation and sensitive amplification-free detection of Ebola virus

Extraction of High Quality RNA from Cannabis sativa Bast Fibres: A Vademecum for Molecular Biologists

The quality is in the eye of the beholder: The perspective of FTIR and UV resonant Raman spectroscopies on extracted nucleic acids

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