scholarly journals Modification of the Acid Guanidinium Thiocyanate-Phenol-Chloroform Method for Nuclear RNA Isolation

BioTechniques ◽  
1996 ◽  
Vol 21 (2) ◽  
pp. 236-237 ◽  
Author(s):  
Katerina Vareli ◽  
Maria Frangou-Lazaridis
Keyword(s):  
Rna Isolation ◽  
Guanidinium Thiocyanate ◽  
Nuclear Rna

Nuclear RNA Isolation and Sequencing

2021 ◽  
pp. 75-83
Author(s):  
Navroop K. Dhaliwal ◽  
Jennifer A. Mitchell
Keyword(s):  
Rna Isolation ◽  
Nuclear Rna

scholarly journals MAVRICS: A Robust and Safe Magnetic Nanoparticle based RNA Extraction Method Compatible with Phenol-chloroform Inactivated Infectious Samples v2

2021 ◽  
Author(s):  
Mo Li ◽  
Gerardo Ramos-Mandujano
Keyword(s):  
Extraction Method ◽  
Magnetic Nanoparticle ◽  
Rna Extraction ◽  
Rna Isolation ◽  
Emerging Pathogens ◽  
Nucleic Acid Isolation ◽  
Guanidinium Thiocyanate ◽  
Biology Laboratory ◽  
Alternative Solutions ◽  
Basic Equipment

Diagnosis and surveillance of emerging pathogens such as SARS-CoV-2 depend on nucleic acid isolation from clinical and environmental samples. Under normal circumstances, samples would be processed using commercial proprietary reagents in Biosafety 2 (BSL-2) or higher facilities. A pandemic at the scale of COVID-19 has caused a global shortage of proprietary reagents and BSL-2 laboratories to safely perform testing. Therefore, alternative solutions are urgently needed to address these challenges. We developed an open-source method called Magneticnanoparticle-Aided Viral RNA Isolation of Contagious Samples (MAVRICS) that is built upon reagents that are either readily available or can be synthesized in any molecular biology laboratory with basic equipment. Unlike conventional methods, MAVRICS works directly in samples inactivated in acid guanidinium thiocyanate-phenol-chloroform (e.g., TRIzol), thus allowing infectious samples to be handled safely without biocontainment facilities.

scholarly journals RNA Isolation of Dengue Virus Type 2 with Different Precipitation Solvents : Methanol, Chloroform, and 2-Isopropanol.

2018 ◽  
Vol 3 (1) ◽  
pp. 52
Author(s):  
Yovilianda Maulitiva Untoro ◽  
Teguh Hari Sucipto ◽  
Harsasi Setyawati ◽  
Siti Churrotin ◽  
Ilham Harlan Amarullah ◽  
...  
Keyword(s):  
Aqueous Solution ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Virus Type ◽  
Aedes Albopictus ◽  
Rna Isolation ◽  
Guanidinium Thiocyanate ◽  
The World ◽  
Dengue Virus Type 2

Dengue virus distributed in tropical and subtropical regions in the world. DENV viruses are transmitted between humans primarily by Aedes aegypti and Aedes albopictus mosquitoes and are endemic in most areas in which the vectors occur. Four serotypes of dengue virus are DENV-1, DENV-2, DENV-3 and DENV-4. DENV-2 is comprised of six genotypes. Ethanol precipitation is a commonly used technique for concentrating and de-salting nucleic acids (DNA or RNA) preparations in aqueous solution. RNA isolation by combining Guanidinium thiocyanate and phenol reported has been reported. In this report, we investigated RNA isolation from DENV-2 using QIAamp Mini Kit with 2-Isopropanol, Methanol, Chloroform precipitation solvent. Electrophoregram showed DNA band as  the result of RNA isolation with methanol and 2-isopropanol are produced quite well. Dna band of the of RNA isolation with chloroform solvent has the lowest intensity than methanol and 2-isopropanol. This study showed that methanol and 2-isopropanol  can used as precipitation solvent for isolating RNA.

Nuclear RNA Isolation and Sequencing

2016 ◽  
pp. 63-71 ◽  
Author(s):  
Navroop K. Dhaliwal ◽  
Jennifer A. Mitchell
Keyword(s):  
Rna Isolation ◽  
Nuclear Rna

RNA Isolation Protocol from Cells and Tissues

2020 ◽  
pp. 7-14
Keyword(s):  
Rna Structure ◽  
Hydrophobic Interactions ◽  
Secretory Granules ◽  
Rna Isolation ◽  
Tertiary Structures ◽  
Guanidinium Thiocyanate ◽  
A Cell ◽  
High Concentrations ◽  
Protein Denaturant ◽  
Chaotropic Agents

The preparation of intact ribonucleic acid is difficult because of the action of nucleases, which are liberated upon tissue homogenisation. In many cells, high concentrations of the ribonucleases are reserved in the secretory granules and upon disruption of the cell, they get mixed with the RNA and lead to its degradation. Guanidinium chloride and thiocyanate are potent chaotropic agents that reduce hydrophobic interactions and disrupt protein tertiary structures, disassociate proteinnucleic acid complexes and disintegrate cellular structures. Guanidinium thiocyanate is especially strong protein denaturant because both the cation and anion disrupt the hydrophobic bonds between the amino acid side chains. RNA usually binds to proteins within a cell and this agent disassociates the nucleoprotein complex, without disrupting RNA structure. Thus RNA can be obtained by using these agents, after homogenisation and low-speed centrifugation and precipitated with ethanol. The protocol below explains the stepwise isolation of total RNA from cells and tissues using TRIzol reagent which is the mono-phasic solution of phenol and guanidine thiocyanate.

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