kinasemax 5 labeling kit  (Thermo Fisher)


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    Name:
    KinaseMax 5 End Labeling Kit
    Description:
    The Ambion KinaseMax 5 End Labeling Kit allows the efficient end labeling of DNA or RNA to high specific activity with T4 polynucleotide kinase and gamma 32P ATP or quantitative phosphorylation of 5 ends using unlabeled ATP Includes sufficient reagents for 30 reactions • Label oligonucleotide probes for nuclease protection assays and blot hybridizations• Label primers for northerns and quantitative PCR• Obtain up to 3 fold greater yields than with standard kinase buffers• Reagents included for forward and dephosphorylation reactionsThe kit includes a kinase reaction buffer that exceeds the performance of the standard forward reaction buffers recommended in Molecular Cloning A Laboratory Manual Sambrook et al 1989 and Current Protocols in Molecular Biology John Wiley Sons Inc Ausubel FM et al editors 1994 Crude 7000 Ci mmol gamma 32P ATP is compatible with this kit End labeled probes are more stable than internally labeled probes and can routinely be used in assays for two to four weeks Rapid Phenol Free Phosphatase Removal StepInactivating the Calf Intestinal Phosphatase CIP after the dephosphorylation reaction is a time consuming and cumbersome procedure that usually requires phenol extraction and ethanol precipitation typically resulting in sample loss The KinaseMax Kit now includes a novel Phosphatase Removal Reagent for quick and complete removal of CIP After the dephosphorylation step the reagent is added to the reaction incubated at room temperature for 2 minutes The tube is then spun for a minute in the microfuge and the supernatant transferred to a new tube for the kinasing reaction Accessory Products The NucAway Spin Columns SKU AM10070 are available for rapid purification of end labeled probes
    Catalog Number:
    am1520
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Labeling|Northern Blotting|Nuclease Protection Assays|Nucleic Acid Labeling & Oligo Synthesis|Oligo Labeling|RNA Labeling|Southern Blotting|Nucleic Acid Gel Electrophoresis & Blotting
    Category:
    Kits and Assays
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    Structured Review

    Thermo Fisher kinasemax 5 labeling kit
    The Ambion KinaseMax 5 End Labeling Kit allows the efficient end labeling of DNA or RNA to high specific activity with T4 polynucleotide kinase and gamma 32P ATP or quantitative phosphorylation of 5 ends using unlabeled ATP Includes sufficient reagents for 30 reactions • Label oligonucleotide probes for nuclease protection assays and blot hybridizations• Label primers for northerns and quantitative PCR• Obtain up to 3 fold greater yields than with standard kinase buffers• Reagents included for forward and dephosphorylation reactionsThe kit includes a kinase reaction buffer that exceeds the performance of the standard forward reaction buffers recommended in Molecular Cloning A Laboratory Manual Sambrook et al 1989 and Current Protocols in Molecular Biology John Wiley Sons Inc Ausubel FM et al editors 1994 Crude 7000 Ci mmol gamma 32P ATP is compatible with this kit End labeled probes are more stable than internally labeled probes and can routinely be used in assays for two to four weeks Rapid Phenol Free Phosphatase Removal StepInactivating the Calf Intestinal Phosphatase CIP after the dephosphorylation reaction is a time consuming and cumbersome procedure that usually requires phenol extraction and ethanol precipitation typically resulting in sample loss The KinaseMax Kit now includes a novel Phosphatase Removal Reagent for quick and complete removal of CIP After the dephosphorylation step the reagent is added to the reaction incubated at room temperature for 2 minutes The tube is then spun for a minute in the microfuge and the supernatant transferred to a new tube for the kinasing reaction Accessory Products The NucAway Spin Columns SKU AM10070 are available for rapid purification of end labeled probes
    https://www.bioz.com/result/kinasemax 5 labeling kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kinasemax 5 labeling kit - by Bioz Stars, 2020-04
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    Related Articles

    Clone Assay:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: All the fragments were cloned in the pGEM-T Easy sub-cloning vector (Promega) and their sequences were verified. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Centrifugation:

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: RNA was recovered by centrifugation, resuspended in nuclease-free water, and its concentration determined by nanodrop reading. .. RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520).

    Amplification:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: Genomic fragments containing pri-miRNA156a and pri-miRNA166b were amplified using primer sets pri-156a-T7 and pri-156a-R and pri-166b-T7 and pri-166b-R, respectively. .. To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates.

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: For bait DNA preparation, promoter fragments P1, P2, and P3 of prom:St GH3.6 were PCR amplified from potato genomic DNA, and promoter fragment P4 of prom:At GH3.5 was PCR amplified from Arabidopsis genomic DNA. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Synthesized:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: RNA substrates were synthesized using the Megascript T7 kit (Ambion, AM1334). .. To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates.

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit. .. After extensive washing with wash buffer at room temperature, the membrane was air-dried for autoradiography at −80 °C.

    Autoradiography:

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit. .. After extensive washing with wash buffer at room temperature, the membrane was air-dried for autoradiography at −80 °C.

    Article Title: Identification of spermidine binding site in T box riboswitch antiterminator RNA
    Article Snippet: For all probing studies, AM1A was labeled with 32 P using Ambion® KinaseMax™ 5′ end-labeling kit from Life Technologies (Grand Island, NY, USA). .. The cleavage products were separated on a 20% denaturing polyacrylamide gel (19:1 acrylamide : bis-acrylamide), followed by autoradiography.

    Quantitative RT-PCR:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: Paragraph title: Northern blot and real time RT–PCR ... End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Electrophoresis:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: For northern blot analysis, 10 µg total RNA from each sample was separated by electrophoresis on a 1% glyoxal agarose gel using the NorthernMax™-Gly Glyoxal-Based System (Ambion). .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Incubation:

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: .. Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.). ..

    Article Title: mRNA Decay during Herpes Simplex Virus (HSV) Infections: Mutations That Affect Translation of an mRNA Influence the Sites at Which It Is Cleaved by the HSV Virion Host Shutoff (Vhs) Protein
    Article Snippet: The mRNAs were dephosphorylated with calf intestine alkaline phosphatase according to procedures described for the KinaseMax 5′-end-labeling kit (Ambion), after which the phosphatase was removed using phosphatase removal reagent (Ambion), and the mRNAs were precipitated from ethanol. .. The mRNAs were resuspended in a small volume of water and incubated with polynucleotide kinase in the presence of 1 mM ATP to add a 5′ monophosphate.

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: After incubation for 2 h at 37°C, DNase was added and incubation continued for further 15 min, followed by phenol extraction and overnight precipitation in ethanol/sodium acetate/glycogen. .. RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520).

    Article Title: Identification of spermidine binding site in T box riboswitch antiterminator RNA
    Article Snippet: For all probing studies, AM1A was labeled with 32 P using Ambion® KinaseMax™ 5′ end-labeling kit from Life Technologies (Grand Island, NY, USA). .. For probing with RNaseA, the 5′-32 P-AM1A was incubated in 10 mM Tris, pH 7.0, 100 mM KCl, 10 mM MgCl2 with spermidine (0–7 mM) and RNase A (0.02 μg/mL) for 15 min at room temperature.

    Expressing:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: Protein expression was performed using Escherichia coli BL21(DE3) cells as host, followed by Ni-NTA affinity column-based protein purification. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Modification:

    Article Title: mRNA Decay during Herpes Simplex Virus (HSV) Infections: Mutations That Affect Translation of an mRNA Influence the Sites at Which It Is Cleaved by the HSV Virion Host Shutoff (Vhs) Protein
    Article Snippet: Circular forms of the pBK2 and pCITE-RLuc mRNAs were prepared using a modification of the procedure described by Chen and Sarnow ( , ). .. The mRNAs were dephosphorylated with calf intestine alkaline phosphatase according to procedures described for the KinaseMax 5′-end-labeling kit (Ambion), after which the phosphatase was removed using phosphatase removal reagent (Ambion), and the mRNAs were precipitated from ethanol.

    RNA Binding Assay:

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: The blot was then renatured in RNA binding buffer and hybridized to a 5′-end-labeled RNA probe (∼106 cpm/5 ml of hybridization buffer) overnight at ambient temperature. .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit.

    Hybridization:

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: The blot was then renatured in RNA binding buffer and hybridized to a 5′-end-labeled RNA probe (∼106 cpm/5 ml of hybridization buffer) overnight at ambient temperature. .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit.

    Immunoprecipitation:

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: Briefly, GFP or GFP-tagged proteins were immunoprecipitated from H1299 cells as described above, separated on 4–12% SDS-polyacrylamide gel, and transferred to nitrocellulose membrane. .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit.

    Northern Blot:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: Paragraph title: Northern blot and real time RT–PCR ... End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Generated:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: The MAD-3 probe was generated by end-labeling a DNA oligonucleotide containing the sequence 5′-GCCCCTTTGCACTCATAACGTCAGACGCTGGCCTCCAAACACACAGTCATCATAGGGC-3′) and the TNFSF14 probe was generated by end-labeling the DNA oligonucleotide 5′-GGCACCCTCTGAGTTCTCCACGTGTCAGACCCATGTCCAATGCACCACGCTCC-3′. .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Polymerase Chain Reaction:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: The resulting 133- and 216-bp PCR fragments were gel purified and used as the templates to synthesize RNA substrates. .. To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates.

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: For bait DNA preparation, promoter fragments P1, P2, and P3 of prom:St GH3.6 were PCR amplified from potato genomic DNA, and promoter fragment P4 of prom:At GH3.5 was PCR amplified from Arabidopsis genomic DNA. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions. .. PCR was then performed using p27kip1 specific forward and reverse primers (5′-TTCAGACGGTTCCCCAAAT-3′ and 5′-AACGCTTTTAGAGGCAGATCA-3′).

    DNA Labeling:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: The GAPDH probe was generated using the DECAprime™ II Random Priming DNA Labeling Kit (Ambion) according to the manufacturer’s protocol. .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions. .. The p27kip1 probe was generated by RT–PCR. cDNA was generated using the ProSTAR™ Ultra HF RT–PCR System (Stratagene) using total RNA from purified human T cells.

    Binding Assay:

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520). .. Before use in binding and structural studies, RNA was heated in refolding buffer (50 mM Tris at pH 8, 0.1 M NaCl, 0.1 M KCl, 1 mM MgCl2 ) for 3 min at 85°C, followed by 20 min at room temperature, and finally placed on ice.

    Subcloning:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: All the fragments were cloned in the pGEM-T Easy sub-cloning vector (Promega) and their sequences were verified. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Size-exclusion Chromatography:

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.). .. Pre-steady-state kinetics were measured during the first few turnovers ( < 100 sec), while the steady-state kinetics (the initial rate constants) were determined from the initial rate of pre-tRNA cleavage when < 10% of the substrate was cleaved.

    Labeling:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: .. To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates. ..

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: .. Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.). ..

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion). ..

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit. .. After extensive washing with wash buffer at room temperature, the membrane was air-dried for autoradiography at −80 °C.

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: .. RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520). .. Labeled RNA was purified by 8% PAGE.

    Article Title: Identification of spermidine binding site in T box riboswitch antiterminator RNA
    Article Snippet: .. For all probing studies, AM1A was labeled with 32 P using Ambion® KinaseMax™ 5′ end-labeling kit from Life Technologies (Grand Island, NY, USA). .. For probing with RNaseA, the 5′-32 P-AM1A was incubated in 10 mM Tris, pH 7.0, 100 mM KCl, 10 mM MgCl2 with spermidine (0–7 mM) and RNase A (0.02 μg/mL) for 15 min at room temperature.

    Purification:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: The resulting 133- and 216-bp PCR fragments were gel purified and used as the templates to synthesize RNA substrates. .. To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates.

    Article Title: Interactions of Nucleolin and Ribosomal Protein L26 (RPL26) in Translational Control of Human p53 mRNA *
    Article Snippet: .. RNA probes were first synthesized using the MEGAscript high yield transcription kit, labeled at the 5′-end using a KinaseMaxTM 5′-end labeling kit (Ambion), and later purified using the MEGAclear kit. .. After extensive washing with wash buffer at room temperature, the membrane was air-dried for autoradiography at −80 °C.

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: Paragraph title: In vitro RNA synthesis, purification, and labeling ... RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520).

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: Purified human T lymphocytes were stimulated for 3 h with medium or αCD3+αCD28, Act D was added, and total cellular RNA was harvested at 0, 45, 90 and 120 min time points. .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Protein Purification:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: Protein expression was performed using Escherichia coli BL21(DE3) cells as host, followed by Ni-NTA affinity column-based protein purification. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Sequencing:

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: The RNase P substrates, wild-type S. cerevisiae pre-tRNATyr with the intervening sequence, 5′ 12-nt leader, and with or without the 3′-UUUUU trailer [designated as (+5′, +3′) or (+5′, −3′) substrates, respectively] are prepared by in vitro transcription from linearized plasmids catalyzed by T7 RNA polymerase ( ). .. Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.).

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: Electrophoretic mobility shift assay For St ARF10-6×His protein preparation, the coding sequence of St ARF10 was PCR amplified and cloned into the pET28a+ vector (Novagen). .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: The MAD-3 probe was generated by end-labeling a DNA oligonucleotide containing the sequence 5′-GCCCCTTTGCACTCATAACGTCAGACGCTGGCCTCCAAACACACAGTCATCATAGGGC-3′) and the TNFSF14 probe was generated by end-labeling the DNA oligonucleotide 5′-GGCACCCTCTGAGTTCTCCACGTGTCAGACCCATGTCCAATGCACCACGCTCC-3′. .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Electrophoretic Mobility Shift Assay:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion). ..

    Polyacrylamide Gel Electrophoresis:

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520). .. Labeled RNA was purified by 8% PAGE.

    Activated Clotting Time Assay:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: Purified human T lymphocytes were stimulated for 3 h with medium or αCD3+αCD28, Act D was added, and total cellular RNA was harvested at 0, 45, 90 and 120 min time points. .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    Plasmid Preparation:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: All the fragments were cloned in the pGEM-T Easy sub-cloning vector (Promega) and their sequences were verified. .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion).

    Agarose Gel Electrophoresis:

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: For northern blot analysis, 10 µg total RNA from each sample was separated by electrophoresis on a 1% glyoxal agarose gel using the NorthernMax™-Gly Glyoxal-Based System (Ambion). .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions.

    In Vitro:

    Article Title: The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C]The Helicase and RNaseIIIa Domains of Arabidopsis Dicer-Like1 Modulate Catalytic Parameters during MicroRNA Biogenesis 1 [C] [W]The Helicase and
    Article Snippet: Paragraph title: In Vitro miRNA Processing ... To measure the initial velocity of DCL1 enzymes, RNAs 5′ labeled with γ P32 ATP using the KinaseMax kit (Ambion, AM1520) were used as substrates.

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: The RNase P substrates, wild-type S. cerevisiae pre-tRNATyr with the intervening sequence, 5′ 12-nt leader, and with or without the 3′-UUUUU trailer [designated as (+5′, +3′) or (+5′, −3′) substrates, respectively] are prepared by in vitro transcription from linearized plasmids catalyzed by T7 RNA polymerase ( ). .. Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.).

    Article Title: mRNA Decay during Herpes Simplex Virus (HSV) Infections: Mutations That Affect Translation of an mRNA Influence the Sites at Which It Is Cleaved by the HSV Virion Host Shutoff (Vhs) Protein
    Article Snippet: Initially, linear mRNAs with 5′ triphosphates were prepared by in vitro transcription of linearized pBK2 and pCITE-RLuc using the SP6 and T7 RiboMAX in vitro transcription kits (Promega). .. The mRNAs were dephosphorylated with calf intestine alkaline phosphatase according to procedures described for the KinaseMax 5′-end-labeling kit (Ambion), after which the phosphatase was removed using phosphatase removal reagent (Ambion), and the mRNAs were precipitated from ethanol.

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: Paragraph title: In vitro RNA synthesis, purification, and labeling ... RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520).

    Concentration Assay:

    Article Title: Pre-tRNA turnover catalyzed by the yeast nuclear RNase P holoenzyme is limited by product release
    Article Snippet: Pre-tRNA substrates labeled at the 5′ terminus with 32 P were prepared post-transcriptionally by treating RNA with calf intestinal alkaline phosphatase followed by incubation with [γ-32 P]ATP and T4 polynucleotide kinase (KinaseMax 5′ Labeling Kit; Ambion, Inc.). .. The pre-steady-state and steady-state kinetics catalyzed by yeast nuclear RNase P holoenzyme were characterized under conditions where the substrate concentration is in excess to the enzyme ([S] > [E]) in Buffer A ( ; ).

    Article Title: Caught at its own game: regulatory small RNA inactivated by an inducible transcript mimicking its target
    Article Snippet: RNA was recovered by centrifugation, resuspended in nuclease-free water, and its concentration determined by nanodrop reading. .. RNA (10 pmol) was dephosphorylated and labeled at its 5′ end with [γ−32 P]ATP (3000 Ci/mmol from Perkin-Elmer) using the KinaseMax kit (Ambion AM1520).

    End Labeling:

    Article Title: MiRNA160 is associated with local defense and systemic acquired resistance against Phytophthora infestans infection in potato
    Article Snippet: .. For the electrophoretic mobility shift assay (EMSA), probes were prepared by labeling promoter fragments with γ-32 P-ATP using a KinaseMax End-Labeling kit (Ambion). ..

    Article Title: Identification of spermidine binding site in T box riboswitch antiterminator RNA
    Article Snippet: .. For all probing studies, AM1A was labeled with 32 P using Ambion® KinaseMax™ 5′ end-labeling kit from Life Technologies (Grand Island, NY, USA). .. For probing with RNaseA, the 5′-32 P-AM1A was incubated in 10 mM Tris, pH 7.0, 100 mM KCl, 10 mM MgCl2 with spermidine (0–7 mM) and RNase A (0.02 μg/mL) for 15 min at room temperature.

    Article Title: Genome-wide analysis of mRNA decay in resting and activated primary human T lymphocytes
    Article Snippet: .. End-labeling reactions were performed using a KinaseMax™ 5′ End-Labeling Kit (Ambion) according to the manufacturer’s directions. .. The p27kip1 probe was generated by RT–PCR. cDNA was generated using the ProSTAR™ Ultra HF RT–PCR System (Stratagene) using total RNA from purified human T cells.