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5 × reverse transcriptase reaction buffer  (Promega)

 
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    Promega 5 × reverse transcriptase reaction buffer
    5 × Reverse Transcriptase Reaction Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/reverse+transcriptase+buffer/5%C3%97+reverse+transcriptase+buffer/pm11322488-29-20-24
    Average 90 stars, based on 1 article reviews
    5 × reverse transcriptase reaction buffer - by Bioz Stars, 2026-09
    90/100 stars

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    other:

    Article Title: Stereotyped antibody responses target posttranslationally modified gluten in celiac disease
    Article Snippet: Next, 10 μl of 5× reverse transcription (RT) buffer, 3.5 μl of 100 mM DTT, 4.3 μl of 10 mM dNTP-Mix (Promega), 0.7 μl of 40 U/μl RNasin (Promega), 0.85 μl superscript III (Invitrogen), and 4.5 μl DEPC-treated water were added. cDNA was synthesized at 42°C for 10 minutes, 25°C for 10 minutes, 50°C for 60 minutes, and 94°C for 5 minutes and stored at –20°C.

    Article Title: Effects of combined siRNA-TR and -TERT on telomerase activity and growth of bladder transitional cell cancer BIU-87 cells.
    Article Snippet: The effects of combined RNA interference (RNAi) of human telomerase RNA (hTR) and human telomerase reverse transcriptase (hTERT) genes on telomerase activity in a bladder cancer cell line (BIU-87 cells) were investigated by using gene chip technology in vitro with an attempt to evaluate the role of RNAi in the gene therapy of bladder transitional cell cancer (BTCC).. Three TR-specific double-stranded small interfering RNAs (siRNAs) and three TERT-specific double-stranded siRNAs were designed to target different regions of TR and TERT mRNA.. The phTR-siRNA, phTERT-siRNA, and the combination of both plasmids phTR+phTERT-siRNA were transfected into BIU-87 cells.

    Article Title: Phosphatidylcholine causes adipocyte-specific lipolysis and apoptosis in adipose and muscle tissues
    Article Snippet: RNAs were transcribed to cDNA at 40°C for 60 min in a 20-μL cocktail containing 5× reverse transcriptase (RT) buffer, 15 mM dNTPs (300 U), Maloney murine leukemia virus reverse transcriptase (MMLV-RT) (Promega, Madison, WI, USA) and 100 pmol oligo-dT primer. cDNA concentration was estimated by the quantitative RT-PCR method using 2× iQTM SYBR Green Supermix (Bio-Rad, Hercules, CA, USA), to measure the mRNA levels of each gene.

    Reverse Transcription:

    Article Title: Differential response to stress in Ostrea lurida (Carpenter 1864) as measured by gene expression
    Article Snippet: The RNA was combined with 0.25ug of 150 primers in a volume of 74.75uL, incubated at 70°C for 5 minutes in a thermal cycler without a 151 PeerJ PrePrints | https://doi.org/10.7287/peerj.preprints.1595v1 | CC-BY 4.0 Open Access | rec: 18 Dec 2015, publ: 18 Dec 2015 heated lid (PTC-200; MJ Research), and immediately placed on ice. .. Added 25.25uL to each 152 RNA-primer mix of a master mix of 5x Reverse Transcriptase Buffer (Promega), 10mM each of 153 dNTPs (Promega), and M-MMLV Reverse Transcriptase (50U/reactions). .. Samples were 154 incubated at 42°C for 1hr and then at 95°C for 3 minutes in a thermal cycler without a heated lid 155 (PTC-200; MJ Research) and then stored at -20°C.

    Article Title: Differential response to stress in Ostrea lurida as measured by gene expression
    Article Snippet: The RNA was combined with primers (0.25ug) in a volume of 74.75uL, incubated at 70°C for 5 minutes in a thermal cycler without a heated lid (PTC-200; MJ Research), and immediately placed on ice. .. A master mix of 5x Reverse Transcriptase Buffer (1x final concentration; Promega), 10mM each of dNTPs (0.5mM final concentration of each dNTP; Promega), and M-MLV Reverse Transcriptase (50U/reaction) was made and 25.25μL of the mix was added to each sample (final reaction volume 100μL). .. Samples were incubated at 42°C for 1hr, followed by 95°C for 3 minutes in a thermal cycler without a heated lid (PTC-200; MJ Research), and then stored at -20°C.

    Article Title: Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
    Article Snippet: .. Total RNA was extracted from the cells using TRIzol reagent (Gibco-BRL Life Technologies; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and used for cDNA synthesis along with an oligo-dT primer, ImProm-II reverse transcriptase (2 U), 0.5 mM dNTP, 3 mM MgCl 2 , and RNase inhibitor in 5X Reverse Transcriptase Buffer (Promega Co., Madison, WI, USA). cDNA was synthesized at 25°C for 5 min and 42°C for 60 min. PCR was performed with the incubation mixture [2 μ l cDNA, 4 μ M 5′ and 3′ specific primers , 10X buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 25 mM MgCl 2 , 0.1% Triton X-100, 250 μ M dNTP, and 1 U Taq polymerase (TaKaRa Bio Inc., Shiga, Japan)] under the following conditions: 30 sec at 94°C (denaturation), 30 sec at 60°C (annealing), 1 min for extension, and a final extension for 10 min at the end of 35 cycles. .. The band intensities were quantified by densitometric analysis (ChemiDoc MP Imaging System; Bio-Rad Laboratories) and were expressed relative to the intensity of the GAPDH band.

    Article Title: Differential response to stress in Ostrea lurida as measured by gene expression
    Article Snippet: The RNA was combined with primers (0.25 μg) in a volume of 74.75 uL, incubated at 70 °C for 5 min in a thermal cycler without a heated lid (PTC-200; MJ Research), and immediately placed on ice. .. A master mix of 5× Reverse Transcriptase Buffer (1× final concentration; Promega), 10 mM each of dNTPs (0.5 mM final concentration of each dNTP; Promega, Madison, WI, USA), and M-MLV Reverse Transcriptase (50 U/reaction) was made and 25.25 μL of the mix was added to each sample (final reaction volume 100 μL). .. Samples were incubated at 42 °C for 1 h, followed by 95 °C for 3 min in a thermal cycler without a heated lid (PTC-200; MJ Research, Waltham, MA, USA), and then stored at −20 °C.

    Article Title: A Multiplex Reverse Transcription (RT) High-Fidelity PCR Protocol for the Detection of Six Viruses that Cause Potato Tuber Necrosis
    Article Snippet: Viruses that cause necrotic symptoms in potato tubers can be difficult to distinguish based on symptoms and frequently require multiple molecular tests to identify the pathogen.. In this study, a multiplex RT PCR high fidelity PCR protocol was developed using previously validated primers that could accurately detect six important potato viruses and detect multiple viruses in a single sample simultaneously.. To test the protocol, 53 tubers previously tested using conventional PCR were retested using this multiplex protocol and blindly evaluated.

    Concentration Assay:

    Article Title: Differential response to stress in Ostrea lurida as measured by gene expression
    Article Snippet: The RNA was combined with primers (0.25ug) in a volume of 74.75uL, incubated at 70°C for 5 minutes in a thermal cycler without a heated lid (PTC-200; MJ Research), and immediately placed on ice. .. A master mix of 5x Reverse Transcriptase Buffer (1x final concentration; Promega), 10mM each of dNTPs (0.5mM final concentration of each dNTP; Promega), and M-MLV Reverse Transcriptase (50U/reaction) was made and 25.25μL of the mix was added to each sample (final reaction volume 100μL). .. Samples were incubated at 42°C for 1hr, followed by 95°C for 3 minutes in a thermal cycler without a heated lid (PTC-200; MJ Research), and then stored at -20°C.

    Article Title: Differential response to stress in Ostrea lurida as measured by gene expression
    Article Snippet: The RNA was combined with primers (0.25 μg) in a volume of 74.75 uL, incubated at 70 °C for 5 min in a thermal cycler without a heated lid (PTC-200; MJ Research), and immediately placed on ice. .. A master mix of 5× Reverse Transcriptase Buffer (1× final concentration; Promega), 10 mM each of dNTPs (0.5 mM final concentration of each dNTP; Promega, Madison, WI, USA), and M-MLV Reverse Transcriptase (50 U/reaction) was made and 25.25 μL of the mix was added to each sample (final reaction volume 100 μL). .. Samples were incubated at 42 °C for 1 h, followed by 95 °C for 3 min in a thermal cycler without a heated lid (PTC-200; MJ Research, Waltham, MA, USA), and then stored at −20 °C.

    cDNA Synthesis:

    Article Title: Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
    Article Snippet: .. Total RNA was extracted from the cells using TRIzol reagent (Gibco-BRL Life Technologies; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and used for cDNA synthesis along with an oligo-dT primer, ImProm-II reverse transcriptase (2 U), 0.5 mM dNTP, 3 mM MgCl 2 , and RNase inhibitor in 5X Reverse Transcriptase Buffer (Promega Co., Madison, WI, USA). cDNA was synthesized at 25°C for 5 min and 42°C for 60 min. PCR was performed with the incubation mixture [2 μ l cDNA, 4 μ M 5′ and 3′ specific primers , 10X buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 25 mM MgCl 2 , 0.1% Triton X-100, 250 μ M dNTP, and 1 U Taq polymerase (TaKaRa Bio Inc., Shiga, Japan)] under the following conditions: 30 sec at 94°C (denaturation), 30 sec at 60°C (annealing), 1 min for extension, and a final extension for 10 min at the end of 35 cycles. .. The band intensities were quantified by densitometric analysis (ChemiDoc MP Imaging System; Bio-Rad Laboratories) and were expressed relative to the intensity of the GAPDH band.

    Synthesized:

    Article Title: Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
    Article Snippet: .. Total RNA was extracted from the cells using TRIzol reagent (Gibco-BRL Life Technologies; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and used for cDNA synthesis along with an oligo-dT primer, ImProm-II reverse transcriptase (2 U), 0.5 mM dNTP, 3 mM MgCl 2 , and RNase inhibitor in 5X Reverse Transcriptase Buffer (Promega Co., Madison, WI, USA). cDNA was synthesized at 25°C for 5 min and 42°C for 60 min. PCR was performed with the incubation mixture [2 μ l cDNA, 4 μ M 5′ and 3′ specific primers , 10X buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 25 mM MgCl 2 , 0.1% Triton X-100, 250 μ M dNTP, and 1 U Taq polymerase (TaKaRa Bio Inc., Shiga, Japan)] under the following conditions: 30 sec at 94°C (denaturation), 30 sec at 60°C (annealing), 1 min for extension, and a final extension for 10 min at the end of 35 cycles. .. The band intensities were quantified by densitometric analysis (ChemiDoc MP Imaging System; Bio-Rad Laboratories) and were expressed relative to the intensity of the GAPDH band.

    Polymerase Chain Reaction:

    Article Title: Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
    Article Snippet: .. Total RNA was extracted from the cells using TRIzol reagent (Gibco-BRL Life Technologies; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and used for cDNA synthesis along with an oligo-dT primer, ImProm-II reverse transcriptase (2 U), 0.5 mM dNTP, 3 mM MgCl 2 , and RNase inhibitor in 5X Reverse Transcriptase Buffer (Promega Co., Madison, WI, USA). cDNA was synthesized at 25°C for 5 min and 42°C for 60 min. PCR was performed with the incubation mixture [2 μ l cDNA, 4 μ M 5′ and 3′ specific primers , 10X buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 25 mM MgCl 2 , 0.1% Triton X-100, 250 μ M dNTP, and 1 U Taq polymerase (TaKaRa Bio Inc., Shiga, Japan)] under the following conditions: 30 sec at 94°C (denaturation), 30 sec at 60°C (annealing), 1 min for extension, and a final extension for 10 min at the end of 35 cycles. .. The band intensities were quantified by densitometric analysis (ChemiDoc MP Imaging System; Bio-Rad Laboratories) and were expressed relative to the intensity of the GAPDH band.

    Incubation:

    Article Title: Anti-inflammatory and antioxidant effects of MOK, a polyherbal extract, on lipopolysaccharide-stimulated RAW 264.7 macrophages
    Article Snippet: .. Total RNA was extracted from the cells using TRIzol reagent (Gibco-BRL Life Technologies; Thermo Fisher Scientific, Inc., Waltham, MA, USA) and used for cDNA synthesis along with an oligo-dT primer, ImProm-II reverse transcriptase (2 U), 0.5 mM dNTP, 3 mM MgCl 2 , and RNase inhibitor in 5X Reverse Transcriptase Buffer (Promega Co., Madison, WI, USA). cDNA was synthesized at 25°C for 5 min and 42°C for 60 min. PCR was performed with the incubation mixture [2 μ l cDNA, 4 μ M 5′ and 3′ specific primers , 10X buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl, 25 mM MgCl 2 , 0.1% Triton X-100, 250 μ M dNTP, and 1 U Taq polymerase (TaKaRa Bio Inc., Shiga, Japan)] under the following conditions: 30 sec at 94°C (denaturation), 30 sec at 60°C (annealing), 1 min for extension, and a final extension for 10 min at the end of 35 cycles. .. The band intensities were quantified by densitometric analysis (ChemiDoc MP Imaging System; Bio-Rad Laboratories) and were expressed relative to the intensity of the GAPDH band.



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