The Experts below are selected from a list of 306 Experts worldwide ranked by ideXlab platform
Nicoletta Sacchi - One of the best experts on this subject based on the ideXlab platform.
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the single step Method of rna isolation by acid guanidinium thiocyanate phenol chloroform extraction twenty something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:The Single-Step Method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on
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the single step Method of rna isolation by acid guanidinium thiocyanate phenol chloroform extraction twenty something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:Since its introduction, the 'Single-Step' Method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the Method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years.
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The Single-Step Method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:Since its introduction, the 'Single-Step' Method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the Method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years.
Piotr Chomczynski - One of the best experts on this subject based on the ideXlab platform.
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eLS - Single-Step Method of Total RNA Isolation by Guanidine–Phenol Extraction
eLS, 2013Co-Authors: Piotr Chomczynski, William Wilfinger, Karol MackeyAbstract:The original Single-Step Method is the first procedure to isolate purified total ribonucleic acid (RNA) from a variety of sources including tissues and cells from human, animal, plant, yeast, bacterial and viral origins, without the requirement of high-speed ultracentrifugation. The Method is based on liquid-phase separation resulting in sequestration of pure RNA into the aqueous phase. RNA is precipitated from the aqueous phase, dissolved, reprecipitated and washed with alcohol before the final solubilisation step. The entire procedure can be completed in less than 4 h and it provides RNA that is suitable for many sensitive downstream applications such as RNase protection assays, northern blotting, sequencing studies and reverse transcription-polymerase chain reaction. This pioneering Methodology has served as the impetus for the development of newer and improved RNA extraction Methodology that now enable investigators to extract and purify RNA in less than 60 min. Key Concepts: Enzymes within living cells rapidly degrade RNA after a tissue sample is removed from the donor. To prevent RNA degradation, tissue samples that cannot be immediately processed must be rapidly frozen with dry ice or liquid nitrogen and stored frozen at −80 °C until the RNA can be extracted. At the time of RNA extraction, frozen cells or tissues must be immersed in the denaturing solution and rapidly homogenised before the tissue thaws in order to inactivate RNAse and avoid RNA degradation. The quality of the recovered RNA is dependent on a delicate balance of salt concentration and optimal pH. Overloading the extraction solution with too much tissue or diluting the denaturing solution beyond what is specified in the protocol will impact the quality of the resulting RNA. The extraction of RNA from samples that have a high buffering capacity, such as blood, plasma or tissue culture medium, requires greater care in order to maintain optimal salt balance and pH control. Overdrying of the RNA pellets will impede RNA solubilisation. RNA should be solubilised at a concentration that will be appropriate for meaningful spectrophotometric quantitation as well as subsequent downstream molecular biology applications. Enzymes that are involved in RNA degradation are ubiquitous and special care must be taken to avoid RNAse contamination during RNA solubilisation and storage. Keywords: ribonucleic acid; Single-Step Method; liquid-phase separation; total RNA isolation
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RNAzol ® RT: a new Single-Step Method for isolation of RNA
Nature Methods, 2010Co-Authors: Piotr Chomczynski, William Wilfinger, Amy Kennedy, Michal Rymaszewski, Karol MackeyAbstract:The Single-Step RNA isolation with RNAzol® RT yields RNA ready for RT-PCR without additional purification or DNase treatment. Previous Single-Step Methods required additional steps to remove DNA contamination before the isolated RNA was used in RT-PCR. RNAzol RT simplifies the Single-Step Method and adds to its versatility and effectiveness. RNAzol RT isolates pure and undegraded RNA with protocols yielding either a single fraction containing total RNA or two separate fractions containing mRNA and small RNA.
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the single step Method of rna isolation by acid guanidinium thiocyanate phenol chloroform extraction twenty something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:The Single-Step Method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on
-
the single step Method of rna isolation by acid guanidinium thiocyanate phenol chloroform extraction twenty something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:Since its introduction, the 'Single-Step' Method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the Method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years.
-
The Single-Step Method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction: twenty-something years on
Nature Protocols, 2006Co-Authors: Piotr Chomczynski, Nicoletta SacchiAbstract:Since its introduction, the 'Single-Step' Method has become widely used for isolating total RNA from biological samples of different sources. The principle at the basis of the Method is that RNA is separated from DNA after extraction with an acidic solution containing guanidinium thiocyanate, sodium acetate, phenol and chloroform, followed by centrifugation. Under acidic conditions, total RNA remains in the upper aqueous phase, while most of DNA and proteins remain either in the interphase or in the lower organic phase. Total RNA is then recovered by precipitation with isopropanol and can be used for several applications. The original protocol, enabling the isolation of RNA from cells and tissues in less than 4 hours, greatly advanced the analysis of gene expression in plant and animal models as well as in pathological samples, as demonstrated by the overwhelming number of citations the paper gained over 20 years.
Karol Mackey - One of the best experts on this subject based on the ideXlab platform.
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eLS - Single-Step Method of Total RNA Isolation by Guanidine–Phenol Extraction
eLS, 2013Co-Authors: Piotr Chomczynski, William Wilfinger, Karol MackeyAbstract:The original Single-Step Method is the first procedure to isolate purified total ribonucleic acid (RNA) from a variety of sources including tissues and cells from human, animal, plant, yeast, bacterial and viral origins, without the requirement of high-speed ultracentrifugation. The Method is based on liquid-phase separation resulting in sequestration of pure RNA into the aqueous phase. RNA is precipitated from the aqueous phase, dissolved, reprecipitated and washed with alcohol before the final solubilisation step. The entire procedure can be completed in less than 4 h and it provides RNA that is suitable for many sensitive downstream applications such as RNase protection assays, northern blotting, sequencing studies and reverse transcription-polymerase chain reaction. This pioneering Methodology has served as the impetus for the development of newer and improved RNA extraction Methodology that now enable investigators to extract and purify RNA in less than 60 min. Key Concepts: Enzymes within living cells rapidly degrade RNA after a tissue sample is removed from the donor. To prevent RNA degradation, tissue samples that cannot be immediately processed must be rapidly frozen with dry ice or liquid nitrogen and stored frozen at −80 °C until the RNA can be extracted. At the time of RNA extraction, frozen cells or tissues must be immersed in the denaturing solution and rapidly homogenised before the tissue thaws in order to inactivate RNAse and avoid RNA degradation. The quality of the recovered RNA is dependent on a delicate balance of salt concentration and optimal pH. Overloading the extraction solution with too much tissue or diluting the denaturing solution beyond what is specified in the protocol will impact the quality of the resulting RNA. The extraction of RNA from samples that have a high buffering capacity, such as blood, plasma or tissue culture medium, requires greater care in order to maintain optimal salt balance and pH control. Overdrying of the RNA pellets will impede RNA solubilisation. RNA should be solubilised at a concentration that will be appropriate for meaningful spectrophotometric quantitation as well as subsequent downstream molecular biology applications. Enzymes that are involved in RNA degradation are ubiquitous and special care must be taken to avoid RNAse contamination during RNA solubilisation and storage. Keywords: ribonucleic acid; Single-Step Method; liquid-phase separation; total RNA isolation
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RNAzol ® RT: a new Single-Step Method for isolation of RNA
Nature Methods, 2010Co-Authors: Piotr Chomczynski, William Wilfinger, Amy Kennedy, Michal Rymaszewski, Karol MackeyAbstract:The Single-Step RNA isolation with RNAzol® RT yields RNA ready for RT-PCR without additional purification or DNase treatment. Previous Single-Step Methods required additional steps to remove DNA contamination before the isolated RNA was used in RT-PCR. RNAzol RT simplifies the Single-Step Method and adds to its versatility and effectiveness. RNAzol RT isolates pure and undegraded RNA with protocols yielding either a single fraction containing total RNA or two separate fractions containing mRNA and small RNA.
Nur Raidah Salim - One of the best experts on this subject based on the ideXlab platform.
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Convergence of interval symmetric Single-Step Method for simultaneous inclusion of real polynomial zeros
2020Co-Authors: Nur Raidah SalimAbstract:The purpose of this thesis is to find the inclusion of polynomial zeros by using interval analysis approach. We will focus on interval Single-Step Method in order to gain the fastest speed of convergence for bounding simple polynomial zeros simultaneously. Firstly, we will generally describe on some basic mathematical background on interval analysis approach. Then, we will briefly discuss the procedure given in the literature which has been proved by other researchers. We present some information on interval Single-Step IS Method together with the algorithm and the analysis on the rate of convergence. In order to improve IS Method, we made several modifications using interval analysis approaches whereby it has been proved that these procedures not only including intervals for roots, but also convergent under a few assumptions. We have new modification namely ISS, IZSS and IZMSS Methods which are describe precisely in this thesis. The processing time (CPU) of the algorithm of the modified Methods may be done using Matlab 2007a associated with Intlab. Nevertheless, we will also present the theoretical analyses of the convergence rate of the modified procedure. This thesis will cover the algorithms, theoretical analysis and numerical results for each modification. Based on the analysis that has been done, we finally found the rate of convergence for ISS is at least 9, for IZSS is at least 13 and for IMZSS is at least 16 while the rate of convergence of IS is at least 2(1+r)˃3. Finally, we conclude our thesis by comparing all the factors needed in a table and we give some possible extensions for future works.
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On the Convergence Rate of Symmetric Single-Step Method ISS for Simultaneous Bounding Polynomial Zeros
2020Co-Authors: Nur Raidah Salim, Mansor Monsi, Malik Abu Hassan, Wah June LeongAbstract:A new modified Method ISS for the simultaneous bounding all the zeros of a polynomial is formulated in this paper. The efficiency of this Method is measured on the CPU times and the number of iterations after satisfying the convergence criteria where the results are obtained using five tested polynomials. The R-order of convergence of this Method is at least 9.
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On the performances of IMZSS2 Method for bounding polynomial zeros simultaneously
2015 International Conference on Research and Education in Mathematics (ICREM7), 2015Co-Authors: Nur Raidah Salim, Mansor Monsi, Nasruddin HassanAbstract:This paper describes the extension of the interval symmetric Single-Step Method IZSS2, namely the interval midpoint symmetric Single-Step IMZSS2 Method which performs a forward-backward-forward step. The algorithm IMZSS2 introduced new reusable correctors where we always update the midpoints of the intervals at every step of the Method. We will display the numerical results comparing the CPU times and number of iterations of both Methods. The results show that the IMZSS2 Method performs better both in CPU times and number of iterations as can be seen in the accompanied figures.
Michio Kondo - One of the best experts on this subject based on the ideXlab platform.
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Single step Method to deposit Si quantum dot films using H2 + SiH4 VHF discharges and electron mobility in a Si quantum dot solar cell
Surface & Coatings Technology, 2007Co-Authors: Masaharu Shiratani, Kazunori Koga, Soichiro Ando, Toshihisa Inoue, Yukio Watanabe, Shota Nunomura, Michio KondoAbstract:Abstract To fabricate quantum dot solar cells of a high efficiency at a low cost, here we propose a Si quantum dot film which is composed of Si nano-crystallites embedded into a-Si:H and a single step Method to deposit such Si quantum dot films using H 2 + SiH 4 VHF discharges. For the Method, Si nano-crystallites of a small size dispersion and radicals produced in the discharges are co-deposited on a substrate to form Si quantum dot films. Using the Method, we realized a volume fraction of dots in films of 0.3–70%. Photo- and dark-conductivity of films are in a range of 10 − 11 –10 − 9 and 10 − 5 –10 − 4 S/cm, respectively. We have examined effects of the size dispersion of nano-crystallites on electron mobility in a quantum dot Si solar cell by a simulation. The electron mobility in films for a size dispersion of 0.37 nm is about 50% of that in films for no size dispersion.
