Improved method for isolating high-quality RNA from cells exposed to metal chloride solutions and clay mineral suspensions

Shelley Haydel (Inventor)

Research output: Patent

Abstract

Microarrays and high-throughput transcriptomic analyses require high quality RNA samples for accurate results. Extracting high quality RNA is notoriously difficult and can be further complicated by high clay or metal ion concentrations. Nucleic acids are difficult to separate from clay particles and metal ions may bind irreversibly to RNA and cause degradation. Additionally, lengthy wash steps are typically required for RNA isolation and purification, which may alter or degrade RNA populations, and are therefore undesirable for transcriptomic experiments. Researchers at the Biodesign Institute of Arizona State University have developed an extraction buffer and protocol for functional, rapid, and efficient isolation of total microbial RNA from samples containing high concentrations of aqueous metal chlorides or clay. This method has been demonstrated to yield approximately 80-200 g of high quality RNA with limited genomic DNA contamination in less than 90 minutes from approximately 1x109 E. coli cells. This optimized protocol allows for the efficient extraction of ribonucleic acids from cell pellets containing high concentrations of aqueous transition metal chlorides as well as from clay mixtures. Potential Applications High-quality microbial RNA extraction Transcriptomic analyses Benefits and Advantages Efficient and high-yield extraction of intact RNA The entire protocol takes less than 90 minutes Does not require lengthy wash steps prior to cell lysis Contamination of genomic DNA is minimized Precipitation of nucleic acids, salts, and surfactants is limited Dowload Original PDF For more information about the inventor(s) and their research, please see Dr. Haydel's directory webpage Dr. Haydel's departmental webpage Dr. Haydel's laboratory webpage
Original languageEnglish (US)
StatePublished - Nov 8 2012

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Clay minerals
Chlorides
Suspensions
Metals
RNA
Nucleic Acids
Metal ions
Contamination
Precipitation (meteorology)
DNA
Microarrays
Surface-Active Agents
Escherichia coli
Transition metals
Purification
Buffers
Salts
Throughput
Degradation
clay

Cite this

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title = "Improved method for isolating high-quality RNA from cells exposed to metal chloride solutions and clay mineral suspensions",
abstract = "Microarrays and high-throughput transcriptomic analyses require high quality RNA samples for accurate results. Extracting high quality RNA is notoriously difficult and can be further complicated by high clay or metal ion concentrations. Nucleic acids are difficult to separate from clay particles and metal ions may bind irreversibly to RNA and cause degradation. Additionally, lengthy wash steps are typically required for RNA isolation and purification, which may alter or degrade RNA populations, and are therefore undesirable for transcriptomic experiments. Researchers at the Biodesign Institute of Arizona State University have developed an extraction buffer and protocol for functional, rapid, and efficient isolation of total microbial RNA from samples containing high concentrations of aqueous metal chlorides or clay. This method has been demonstrated to yield approximately 80-200 g of high quality RNA with limited genomic DNA contamination in less than 90 minutes from approximately 1x109 E. coli cells. This optimized protocol allows for the efficient extraction of ribonucleic acids from cell pellets containing high concentrations of aqueous transition metal chlorides as well as from clay mixtures. Potential Applications High-quality microbial RNA extraction Transcriptomic analyses Benefits and Advantages Efficient and high-yield extraction of intact RNA The entire protocol takes less than 90 minutes Does not require lengthy wash steps prior to cell lysis Contamination of genomic DNA is minimized Precipitation of nucleic acids, salts, and surfactants is limited Dowload Original PDF For more information about the inventor(s) and their research, please see Dr. Haydel's directory webpage Dr. Haydel's departmental webpage Dr. Haydel's laboratory webpage",
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T1 - Improved method for isolating high-quality RNA from cells exposed to metal chloride solutions and clay mineral suspensions

AU - Haydel, Shelley

PY - 2012/11/8

Y1 - 2012/11/8

N2 - Microarrays and high-throughput transcriptomic analyses require high quality RNA samples for accurate results. Extracting high quality RNA is notoriously difficult and can be further complicated by high clay or metal ion concentrations. Nucleic acids are difficult to separate from clay particles and metal ions may bind irreversibly to RNA and cause degradation. Additionally, lengthy wash steps are typically required for RNA isolation and purification, which may alter or degrade RNA populations, and are therefore undesirable for transcriptomic experiments. Researchers at the Biodesign Institute of Arizona State University have developed an extraction buffer and protocol for functional, rapid, and efficient isolation of total microbial RNA from samples containing high concentrations of aqueous metal chlorides or clay. This method has been demonstrated to yield approximately 80-200 g of high quality RNA with limited genomic DNA contamination in less than 90 minutes from approximately 1x109 E. coli cells. This optimized protocol allows for the efficient extraction of ribonucleic acids from cell pellets containing high concentrations of aqueous transition metal chlorides as well as from clay mixtures. Potential Applications High-quality microbial RNA extraction Transcriptomic analyses Benefits and Advantages Efficient and high-yield extraction of intact RNA The entire protocol takes less than 90 minutes Does not require lengthy wash steps prior to cell lysis Contamination of genomic DNA is minimized Precipitation of nucleic acids, salts, and surfactants is limited Dowload Original PDF For more information about the inventor(s) and their research, please see Dr. Haydel's directory webpage Dr. Haydel's departmental webpage Dr. Haydel's laboratory webpage

AB - Microarrays and high-throughput transcriptomic analyses require high quality RNA samples for accurate results. Extracting high quality RNA is notoriously difficult and can be further complicated by high clay or metal ion concentrations. Nucleic acids are difficult to separate from clay particles and metal ions may bind irreversibly to RNA and cause degradation. Additionally, lengthy wash steps are typically required for RNA isolation and purification, which may alter or degrade RNA populations, and are therefore undesirable for transcriptomic experiments. Researchers at the Biodesign Institute of Arizona State University have developed an extraction buffer and protocol for functional, rapid, and efficient isolation of total microbial RNA from samples containing high concentrations of aqueous metal chlorides or clay. This method has been demonstrated to yield approximately 80-200 g of high quality RNA with limited genomic DNA contamination in less than 90 minutes from approximately 1x109 E. coli cells. This optimized protocol allows for the efficient extraction of ribonucleic acids from cell pellets containing high concentrations of aqueous transition metal chlorides as well as from clay mixtures. Potential Applications High-quality microbial RNA extraction Transcriptomic analyses Benefits and Advantages Efficient and high-yield extraction of intact RNA The entire protocol takes less than 90 minutes Does not require lengthy wash steps prior to cell lysis Contamination of genomic DNA is minimized Precipitation of nucleic acids, salts, and surfactants is limited Dowload Original PDF For more information about the inventor(s) and their research, please see Dr. Haydel's directory webpage Dr. Haydel's departmental webpage Dr. Haydel's laboratory webpage

M3 - Patent

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