t4 rna ligase 2  (Thermo Fisher)


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    Name:
    RNaseOUT Recombinant Ribonuclease Inhibitor
    Description:
    RNaseOUT Recombinant Ribonuclease Inhibitor is a potent noncompetitive inhibitor of pancreatic type ribonucleases such as RNase A and is used to avoid RNA degradation in a variety of applications RNaseOUT Recombinant Ribonuclease Inhibitor is an acidic protein with a molecular weight of 52 kDa RNaseOUT inhibits RNase A RNase B and RNase C ApplicationscDNA synthesis RT PCR and in vitro transcription and translationSourcePurified by affinity chromatography from E coli expressing a cloned porcine genePerformance and quality testingSDS PAGE purity endodeoxyribonuclease assay protein concentration specific activity performance evaluated by RT PCRUnit definitionOne unit inhibits 5 ng of RNase A by 50 using cytidine 2 3 cyclic monophosphate cCMP as the substrateUnit reaction conditions100 mM Tris acetate pH 6 5 1 mM EDTA 0 2 mM cCMP 2 µg RNase A in 1 mL from 0 to 10 min at 25°C
    Catalog Number:
    10777019
    Price:
    None
    Applications:
    PCR & Real-Time PCR|Reverse Transcription
    Category:
    Proteins Enzymes Peptides
    Buy from Supplier


    Structured Review

    Thermo Fisher t4 rna ligase 2
    RNaseOUT Recombinant Ribonuclease Inhibitor is a potent noncompetitive inhibitor of pancreatic type ribonucleases such as RNase A and is used to avoid RNA degradation in a variety of applications RNaseOUT Recombinant Ribonuclease Inhibitor is an acidic protein with a molecular weight of 52 kDa RNaseOUT inhibits RNase A RNase B and RNase C ApplicationscDNA synthesis RT PCR and in vitro transcription and translationSourcePurified by affinity chromatography from E coli expressing a cloned porcine genePerformance and quality testingSDS PAGE purity endodeoxyribonuclease assay protein concentration specific activity performance evaluated by RT PCRUnit definitionOne unit inhibits 5 ng of RNase A by 50 using cytidine 2 3 cyclic monophosphate cCMP as the substrateUnit reaction conditions100 mM Tris acetate pH 6 5 1 mM EDTA 0 2 mM cCMP 2 µg RNase A in 1 mL from 0 to 10 min at 25°C
    https://www.bioz.com/result/t4 rna ligase 2/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    t4 rna ligase 2 - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    In Vivo:

    Article Title: The leishmanicidal activity of oleuropein is selectively regulated through inflammation- and oxidative stress-related genes
    Article Snippet: .. Messenger RNA was then reverse-transcribed using a SuperScript II kit (Invitrogen - Molecular Probes™) and oligo-dTs (Promega, WI, USA), and all reactions included the recombinant ribonuclease inhibitor, RNaseOUT™ (Invitrogen). cDNAs of all the in vivo experimental groups were evaluated with real time PCR, performed using an Exicycler 96 thermocycler (Bioneer, Daejeon, Korea), using the Kapa SYBR Fast Universal 2× qPCR Master Mix kit (Kapa Biosystems Ltd., London, UK). .. The specific primers for the interleukin-1α (IL-1α ), interleukin-1β (IL-1β ), interleukin-1 receptor antagonist (IL-1rn ), interleukin-10 (IL-10 ), subunit 2 of interleukin-12 (IL-12β ), interferon-γ (IFN- γ), tumor necrosis factor-α (TNF-α ), transforming growth factor beta-1 (TGF-β1 ), T-box transcription factor (Tbx21 ), trans-acting T-cell-specific transcription factor (GATA3 ), subunit 2 of nuclear factor kappa-B (NF-kB2) , and the glyceraldehyde dehydrogenase of the 3-phosphatase (GAPDH ) gene sequences were designed by Qiagen (QuantiTect Primer Assays; Qiagen, Venlo, Netherlands).

    In Vitro:

    Article Title: Design and application of cotranscriptional non-enzymatic RNA circuits and signal transducers
    Article Snippet: .. In vitro transcription In all, 100 pg to 1000 ng of double-stranded DNA transcription templates was transcribed using 100 U of T7 RNA polymerase (NEB) in 50 µl reactions containing 40 mM Tris–HCl, pH 7.9, 30 mM MgCl2 , 10 mM DTT, 2 mM spermidine, 4 mM ribonucleotide (rNTP) mix and 20 U of the recombinant ribonuclease inhibitor RNaseOUT (Life Technologies). .. Transcription reactions were incubated at 42°C for 1–2 h. After this, transcripts of the circuit components were either (i) used directly for assembly or (ii) subjected to purification before assembly.

    Protease Inhibitor:

    Article Title: miR-205 Expression Promotes Cell Proliferation and Migration of Human Cervical Cancer Cells
    Article Snippet: .. Cell pellet was collected and then re-suspended in an equal volume (w/v) of lysis buffer [FNN0021; Invitrogen; supplemented with 1 mM Phenylmethanesulfonyl fluoride (PMSF, P7626; Sigma-Aldrich), 1 mM Dithiothreitol ( DTT, 495714; Invitrogen), 1% Protease Inhibitor Cocktail (P8340; Sigma-Aldrich) and 200 U/ml RNaseOUT (10777-019; Invitrogen)], incubated for 10 minutes on ice, and lysed by vortexing. ..

    Real-time Polymerase Chain Reaction:

    Article Title: The leishmanicidal activity of oleuropein is selectively regulated through inflammation- and oxidative stress-related genes
    Article Snippet: .. Messenger RNA was then reverse-transcribed using a SuperScript II kit (Invitrogen - Molecular Probes™) and oligo-dTs (Promega, WI, USA), and all reactions included the recombinant ribonuclease inhibitor, RNaseOUT™ (Invitrogen). cDNAs of all the in vivo experimental groups were evaluated with real time PCR, performed using an Exicycler 96 thermocycler (Bioneer, Daejeon, Korea), using the Kapa SYBR Fast Universal 2× qPCR Master Mix kit (Kapa Biosystems Ltd., London, UK). .. The specific primers for the interleukin-1α (IL-1α ), interleukin-1β (IL-1β ), interleukin-1 receptor antagonist (IL-1rn ), interleukin-10 (IL-10 ), subunit 2 of interleukin-12 (IL-12β ), interferon-γ (IFN- γ), tumor necrosis factor-α (TNF-α ), transforming growth factor beta-1 (TGF-β1 ), T-box transcription factor (Tbx21 ), trans-acting T-cell-specific transcription factor (GATA3 ), subunit 2 of nuclear factor kappa-B (NF-kB2) , and the glyceraldehyde dehydrogenase of the 3-phosphatase (GAPDH ) gene sequences were designed by Qiagen (QuantiTect Primer Assays; Qiagen, Venlo, Netherlands).

    Incubation:

    Article Title: An RNA-seq based comparative approach reveals the transcriptome-wide interplay between 3′-to-5′ exoRNases and RNase Y
    Article Snippet: .. RNAs were annealed to the radiolabelled primer (Supplementary Table ) for 30 min at 65 °C and subsequently incubated on ice for 1 min and the RNAs were reverse transcribed using 1U of SuperScript III Reverse Transcriptase (Invitrogen) in the presence of 1X first strand buffer (Invitrogen), 5 mM dithiothreitol (DTT) and 40 U of RNaseOUT Recombinant Ribonuclease Inhibitor (Invitrogen), for 1 h at 55 °C. .. After inhibition of the SuperScript III enzyme by incubation at 70 °C for 15 min, the cDNAs were precipitated with ice-cold 100% ethanol at –20 °C for 1 h and centrifuged at 20,000 × g for 4 °C for 10 min.

    Article Title: miR-205 Expression Promotes Cell Proliferation and Migration of Human Cervical Cancer Cells
    Article Snippet: .. Cell pellet was collected and then re-suspended in an equal volume (w/v) of lysis buffer [FNN0021; Invitrogen; supplemented with 1 mM Phenylmethanesulfonyl fluoride (PMSF, P7626; Sigma-Aldrich), 1 mM Dithiothreitol ( DTT, 495714; Invitrogen), 1% Protease Inhibitor Cocktail (P8340; Sigma-Aldrich) and 200 U/ml RNaseOUT (10777-019; Invitrogen)], incubated for 10 minutes on ice, and lysed by vortexing. ..

    Lysis:

    Article Title: miR-205 Expression Promotes Cell Proliferation and Migration of Human Cervical Cancer Cells
    Article Snippet: .. Cell pellet was collected and then re-suspended in an equal volume (w/v) of lysis buffer [FNN0021; Invitrogen; supplemented with 1 mM Phenylmethanesulfonyl fluoride (PMSF, P7626; Sigma-Aldrich), 1 mM Dithiothreitol ( DTT, 495714; Invitrogen), 1% Protease Inhibitor Cocktail (P8340; Sigma-Aldrich) and 200 U/ml RNaseOUT (10777-019; Invitrogen)], incubated for 10 minutes on ice, and lysed by vortexing. ..

    Recombinant:

    Article Title: The leishmanicidal activity of oleuropein is selectively regulated through inflammation- and oxidative stress-related genes
    Article Snippet: .. Messenger RNA was then reverse-transcribed using a SuperScript II kit (Invitrogen - Molecular Probes™) and oligo-dTs (Promega, WI, USA), and all reactions included the recombinant ribonuclease inhibitor, RNaseOUT™ (Invitrogen). cDNAs of all the in vivo experimental groups were evaluated with real time PCR, performed using an Exicycler 96 thermocycler (Bioneer, Daejeon, Korea), using the Kapa SYBR Fast Universal 2× qPCR Master Mix kit (Kapa Biosystems Ltd., London, UK). .. The specific primers for the interleukin-1α (IL-1α ), interleukin-1β (IL-1β ), interleukin-1 receptor antagonist (IL-1rn ), interleukin-10 (IL-10 ), subunit 2 of interleukin-12 (IL-12β ), interferon-γ (IFN- γ), tumor necrosis factor-α (TNF-α ), transforming growth factor beta-1 (TGF-β1 ), T-box transcription factor (Tbx21 ), trans-acting T-cell-specific transcription factor (GATA3 ), subunit 2 of nuclear factor kappa-B (NF-kB2) , and the glyceraldehyde dehydrogenase of the 3-phosphatase (GAPDH ) gene sequences were designed by Qiagen (QuantiTect Primer Assays; Qiagen, Venlo, Netherlands).

    Article Title: An RNA-seq based comparative approach reveals the transcriptome-wide interplay between 3′-to-5′ exoRNases and RNase Y
    Article Snippet: .. RNAs were annealed to the radiolabelled primer (Supplementary Table ) for 30 min at 65 °C and subsequently incubated on ice for 1 min and the RNAs were reverse transcribed using 1U of SuperScript III Reverse Transcriptase (Invitrogen) in the presence of 1X first strand buffer (Invitrogen), 5 mM dithiothreitol (DTT) and 40 U of RNaseOUT Recombinant Ribonuclease Inhibitor (Invitrogen), for 1 h at 55 °C. .. After inhibition of the SuperScript III enzyme by incubation at 70 °C for 15 min, the cDNAs were precipitated with ice-cold 100% ethanol at –20 °C for 1 h and centrifuged at 20,000 × g for 4 °C for 10 min.

    Article Title: Design and application of cotranscriptional non-enzymatic RNA circuits and signal transducers
    Article Snippet: .. In vitro transcription In all, 100 pg to 1000 ng of double-stranded DNA transcription templates was transcribed using 100 U of T7 RNA polymerase (NEB) in 50 µl reactions containing 40 mM Tris–HCl, pH 7.9, 30 mM MgCl2 , 10 mM DTT, 2 mM spermidine, 4 mM ribonucleotide (rNTP) mix and 20 U of the recombinant ribonuclease inhibitor RNaseOUT (Life Technologies). .. Transcription reactions were incubated at 42°C for 1–2 h. After this, transcripts of the circuit components were either (i) used directly for assembly or (ii) subjected to purification before assembly.

    Article Title: Notch2 Controls Prolactin and Insulin-Like Growth Factor Binding Protein-1 Expression in Decidualizing Human Stromal Cells of Early Pregnancy
    Article Snippet: .. 2 µg RNA were reverse transcribed using 200U of RevertAid H Minus M.MuLV Reverse Transcriptase (Fermentas, St.Leon-Rot, D), 0.2 µg Hexanucleotide Mix (Roche Diagnostics GmbH, Vienna, Austria), 0.5 µl RNaseOUT (Recombinant Ribonuclease Inhibitor, Invitrogen) and 0.5 mM dNTP (Promega, Madison, WI) in a final volume of 20 µl. .. Quantitative real-time PCR was performed on the ABI 7500 Sequence Detection System (Applied Biosystems, Carlsbad, CA) using Taq Man Gene Expression Assays (TaqMan Universal PCR Master Mix, 20x Taq Man Gene Expression Assay Mix for Notch1 (Hs01062014_m1), Notch2 (Hs01050702_m1), Notch3 (Hs01128541_m1), Notch4 (Hs00965889_m1), Jagged1 (Hs01070032_m1), Jagged2 (Hs00171432_m1), DLL1 (Hs00194509_m1), DLL3 (Hs01085096_m1), DLL4 (Hs00184092_m1), HES1 (Hs00172878_m1), PRL (Hs00168730_m1), IGFBP1 (Hs00426285_m1) and TATA-box binding protein (TBP; TaqMan endogenous control)) according to the manufacturer's instructions.

    Article Title: Genome-wide gene expression analysis of anguillid herpesvirus 1
    Article Snippet: .. RNA was extracted according to manufacturer’s protocol, and dissolved in 200 μl RNase-free water (Sigma-Aldrich) with 2 μl RNaseOUT recombinant ribonuclease inhibitor (Invitrogen). .. RNA concentration was checked by spectrophotometry using a Nanodrop ND-1000 (Thermo Fisher Scientific, MA, USA).

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  • 99
    Thermo Fisher t4 rna ligase 10 u µl
    In vitro preparation of circular RNA sponges. ( A ) Schematic overview of the ciRS production procedure—starting with in vitro transcription with 10-fold excess of GMP, followed by DNase digestion to remove template DNA, gel filtration to clear away excess nucleotides (NTP, GMP) derived from the transcription reaction, and ligation of the purified transcript. Considering the activity of the <t>T4</t> RNA ligase, transcript ligation can result in both linear monomers and dimers of the transcript and circularized transcripts (indicated by the dash/double dash or the circle in (B) and (C)). ( B ) In vitro circularisation reaction of two exemplary constructs was analysed by 7% polyacrylamide-urea gel electrophoresis, followed by ethidium bromide staining showing linear and circular transcripts with a circularisation efficiency of ~ 40%–60% and only small amounts of linear dimers detectable. As described in Section 3.1 , the transcripts are characterized by a constant region and four miRNA binding sites but differ in the presence of a double-stranded stem-loop region increasing the circularisation efficiency (construct A: 197 nt, lacking the 11 nt 3′stem-loop-sequence; construct B: 208 nt, with stem-loop). ( C ) Circular and linear isoforms of the RNA sponges (ciRS and liRS) were purified, and quality was verified on analytic 6% and 7% polyacrylamide-urea gels by ethidium bromide staining. While the mobility of linear RNAs remains unchanged compared to the RNA ladder, the mobility of circular RNA appears lower in higher percentage polyacrylamide-urea gels, resulting in a shift of the circular RNA when comparing the polyacrylamide-urea gels with different concentrations.
    T4 Rna Ligase 10 U µl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 rna ligase 10 u µl/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    t4 rna ligase 10 u µl - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher t4 dna ligase
    In vitro preparation of circular RNA sponges. ( A ) Schematic overview of the ciRS production procedure—starting with in vitro transcription with 10-fold excess of GMP, followed by DNase digestion to remove template DNA, gel filtration to clear away excess nucleotides (NTP, GMP) derived from the transcription reaction, and ligation of the purified transcript. Considering the activity of the <t>T4</t> RNA ligase, transcript ligation can result in both linear monomers and dimers of the transcript and circularized transcripts (indicated by the dash/double dash or the circle in (B) and (C)). ( B ) In vitro circularisation reaction of two exemplary constructs was analysed by 7% polyacrylamide-urea gel electrophoresis, followed by ethidium bromide staining showing linear and circular transcripts with a circularisation efficiency of ~ 40%–60% and only small amounts of linear dimers detectable. As described in Section 3.1 , the transcripts are characterized by a constant region and four miRNA binding sites but differ in the presence of a double-stranded stem-loop region increasing the circularisation efficiency (construct A: 197 nt, lacking the 11 nt 3′stem-loop-sequence; construct B: 208 nt, with stem-loop). ( C ) Circular and linear isoforms of the RNA sponges (ciRS and liRS) were purified, and quality was verified on analytic 6% and 7% polyacrylamide-urea gels by ethidium bromide staining. While the mobility of linear RNAs remains unchanged compared to the RNA ladder, the mobility of circular RNA appears lower in higher percentage polyacrylamide-urea gels, resulting in a shift of the circular RNA when comparing the polyacrylamide-urea gels with different concentrations.
    T4 Dna Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 550 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase/product/Thermo Fisher
    Average 99 stars, based on 550 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher t4 dna ligase buffer
    In vitro preparation of circular RNA sponges. ( A ) Schematic overview of the ciRS production procedure—starting with in vitro transcription with 10-fold excess of GMP, followed by DNase digestion to remove template DNA, gel filtration to clear away excess nucleotides (NTP, GMP) derived from the transcription reaction, and ligation of the purified transcript. Considering the activity of the <t>T4</t> RNA ligase, transcript ligation can result in both linear monomers and dimers of the transcript and circularized transcripts (indicated by the dash/double dash or the circle in (B) and (C)). ( B ) In vitro circularisation reaction of two exemplary constructs was analysed by 7% polyacrylamide-urea gel electrophoresis, followed by ethidium bromide staining showing linear and circular transcripts with a circularisation efficiency of ~ 40%–60% and only small amounts of linear dimers detectable. As described in Section 3.1 , the transcripts are characterized by a constant region and four miRNA binding sites but differ in the presence of a double-stranded stem-loop region increasing the circularisation efficiency (construct A: 197 nt, lacking the 11 nt 3′stem-loop-sequence; construct B: 208 nt, with stem-loop). ( C ) Circular and linear isoforms of the RNA sponges (ciRS and liRS) were purified, and quality was verified on analytic 6% and 7% polyacrylamide-urea gels by ethidium bromide staining. While the mobility of linear RNAs remains unchanged compared to the RNA ladder, the mobility of circular RNA appears lower in higher percentage polyacrylamide-urea gels, resulting in a shift of the circular RNA when comparing the polyacrylamide-urea gels with different concentrations.
    T4 Dna Ligase Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase buffer/product/Thermo Fisher
    Average 99 stars, based on 96 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase buffer - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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    In vitro preparation of circular RNA sponges. ( A ) Schematic overview of the ciRS production procedure—starting with in vitro transcription with 10-fold excess of GMP, followed by DNase digestion to remove template DNA, gel filtration to clear away excess nucleotides (NTP, GMP) derived from the transcription reaction, and ligation of the purified transcript. Considering the activity of the T4 RNA ligase, transcript ligation can result in both linear monomers and dimers of the transcript and circularized transcripts (indicated by the dash/double dash or the circle in (B) and (C)). ( B ) In vitro circularisation reaction of two exemplary constructs was analysed by 7% polyacrylamide-urea gel electrophoresis, followed by ethidium bromide staining showing linear and circular transcripts with a circularisation efficiency of ~ 40%–60% and only small amounts of linear dimers detectable. As described in Section 3.1 , the transcripts are characterized by a constant region and four miRNA binding sites but differ in the presence of a double-stranded stem-loop region increasing the circularisation efficiency (construct A: 197 nt, lacking the 11 nt 3′stem-loop-sequence; construct B: 208 nt, with stem-loop). ( C ) Circular and linear isoforms of the RNA sponges (ciRS and liRS) were purified, and quality was verified on analytic 6% and 7% polyacrylamide-urea gels by ethidium bromide staining. While the mobility of linear RNAs remains unchanged compared to the RNA ladder, the mobility of circular RNA appears lower in higher percentage polyacrylamide-urea gels, resulting in a shift of the circular RNA when comparing the polyacrylamide-urea gels with different concentrations.

    Journal: Methods and Protocols

    Article Title: Production and Purification of Artificial Circular RNA Sponges for Application in Molecular Biology and Medicine

    doi: 10.3390/mps3020042

    Figure Lengend Snippet: In vitro preparation of circular RNA sponges. ( A ) Schematic overview of the ciRS production procedure—starting with in vitro transcription with 10-fold excess of GMP, followed by DNase digestion to remove template DNA, gel filtration to clear away excess nucleotides (NTP, GMP) derived from the transcription reaction, and ligation of the purified transcript. Considering the activity of the T4 RNA ligase, transcript ligation can result in both linear monomers and dimers of the transcript and circularized transcripts (indicated by the dash/double dash or the circle in (B) and (C)). ( B ) In vitro circularisation reaction of two exemplary constructs was analysed by 7% polyacrylamide-urea gel electrophoresis, followed by ethidium bromide staining showing linear and circular transcripts with a circularisation efficiency of ~ 40%–60% and only small amounts of linear dimers detectable. As described in Section 3.1 , the transcripts are characterized by a constant region and four miRNA binding sites but differ in the presence of a double-stranded stem-loop region increasing the circularisation efficiency (construct A: 197 nt, lacking the 11 nt 3′stem-loop-sequence; construct B: 208 nt, with stem-loop). ( C ) Circular and linear isoforms of the RNA sponges (ciRS and liRS) were purified, and quality was verified on analytic 6% and 7% polyacrylamide-urea gels by ethidium bromide staining. While the mobility of linear RNAs remains unchanged compared to the RNA ladder, the mobility of circular RNA appears lower in higher percentage polyacrylamide-urea gels, resulting in a shift of the circular RNA when comparing the polyacrylamide-urea gels with different concentrations.

    Article Snippet: T4 RNA Ligase (Thermo Fisher Scientific, Waltham, MA, USA; Cat. no.: EL0021).

    Techniques: In Vitro, Filtration, Derivative Assay, Ligation, Purification, Activity Assay, Construct, Nucleic Acid Electrophoresis, Staining, Binding Assay, Sequencing