antarctic phosphatase reaction buffer  (New England Biolabs)


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    Name:
    Antarctic Phosphatase Reaction Buffer
    Description:
    Antarctic Phosphatase Reaction Buffer 6 0 ml
    Catalog Number:
    b0289s
    Price:
    23
    Size:
    6 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs antarctic phosphatase reaction buffer
    Antarctic Phosphatase Reaction Buffer
    Antarctic Phosphatase Reaction Buffer 6 0 ml
    https://www.bioz.com/result/antarctic phosphatase reaction buffer/product/New England Biolabs
    Average 90 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    antarctic phosphatase reaction buffer - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Amplification:

    Article Title: Aire controls gene expression in the thymic epithelium with ordered stochasticity
    Article Snippet: First the fragmentation of the amplified RNA (aRNA) was performed using the Magnesium RNA Fragmentation Module (NEBNext E6150S). .. First the 5’ end of the aRNA was dephosphorylated by adding 4 µL of a mix containing 2µL of 10X Antarctic Phosphatase Reaction Buffer, 1µL of Antartic phosphatase (5U/µL stock, NEB M0289) and 1µL of RNAseOut to 16µL of each aRNA pool and incubating at 37°C for 30min and 65°C for 5min.

    Polyacrylamide Gel Electrophoresis:

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Substrate RNA oligonucleotides (below) that contained the suspected splice sites as shown by the RNAse H assays, were generated by run-off transcription and purified by 10% PAGE. .. Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs).

    Synthesized:

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: Capped single stranded RNA was synthesized via IVT using the MEGAscript-T7 kit (Life Technologies). .. To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C.

    Electrophoresis:

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C. .. RNA products were examined by electrophoresis on 2% agarose gels.

    Incubation:

    Article Title: Phosphorylation of ASC acts as a molecular switch controlling the formation of speck-like aggregates and inflammasome activity
    Article Snippet: .. Proteins in the cell lysates were precipitated with acetone, incubated in phosphatase reaction mixture containing 250 U/ml of antarctic phosphatase (New England Biolabs) overnight at 37°C, reprecipitated with acetone and dissolved in SDS sample buffer. ..

    Article Title: Contribution of SAM and HD domains to retroviral restriction mediated by human SAMHD1
    Article Snippet: Reactions were initiated by addition of SAMHD1, incubated for 1 h at 37 °C, and terminated by incubation for 10 min at 70 °C. .. The antarctic phosphatase reaction (2 µl, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Article Title: Aire controls gene expression in the thymic epithelium with ordered stochasticity
    Article Snippet: Samples were immediately incubated at 94°C (lid 105°C) for 2 minutes, then immediately transferred onto ice and the reaction was stopped by adding quickly 2µL of 10X RNA Fragmentation Stop Solution. .. First the 5’ end of the aRNA was dephosphorylated by adding 4 µL of a mix containing 2µL of 10X Antarctic Phosphatase Reaction Buffer, 1µL of Antartic phosphatase (5U/µL stock, NEB M0289) and 1µL of RNAseOut to 16µL of each aRNA pool and incubating at 37°C for 30min and 65°C for 5min.

    Article Title: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1
    Article Snippet: Reactions were initiated by addition of SAMHD1, incubated for 1 h at 37°C, and terminated by incubation for 10 min at 70°C. .. The antarctic phosphatase reaction (2 ul, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: Afterward, the template DNA was removed by adding 1 μL TURBO DNase (from the MEGAscript T7 kit) and incubation at 37°C for 15 min. Then the reaction mixture was purified using the RNeasy MinElute cleanup kit (QIAGEN) according to the manufacturer’s instructions. .. Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA
    Article Snippet: Reactions were incubated 4–6 hours at 37°C and treated with 2 uL TURBO DNase for a further 15 minutes at 37°C before being purified on MEGAclear (Ambion) spin columns, the RNA products being eluted in a volume of 100 uL. .. To remove immunogenic 5′ triphosphate moieties from uncapped transcripts, 10 uL of Antarctic Phosphatase reaction buffer and 3 uL of Antarctic Phosphatase (NEB) was added to each prep.

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Each short substrate was incubated with an excess of spliceozyme to generate the cleavage products. .. Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs).

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: .. To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C. .. Synthesized RNA products were then repurified using RNeasy Mini Columns (Qiagen) and quantitated by spectrophotometry and denaturing gel electrophoresis.

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C. .. 3´ adapter ligation was performed in T4 RNA ligase buffer (NEB) with 10 U/µL T4 RNA ligase 2 (KQ) (NEB), 10 μM 3′ adapter (5′ 5rApp-TGGAATTCTCGGGTGCCAAGG-3ddC 3′ (IDT), 1 U/µL RNase OUT, and 15% (w/v) PEG 8000 overnight at 16°C.

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: .. For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm. ..

    Activity Assay:

    Article Title: Contribution of SAM and HD domains to retroviral restriction mediated by human SAMHD1
    Article Snippet: Paragraph title: Assay to determine dNTPase activity of SAMHD1 ... The antarctic phosphatase reaction (2 µl, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Article Title: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1
    Article Snippet: Paragraph title: Assay to determine dNTPase activity of SAMHD1 by thin-liquid chromatography ... The antarctic phosphatase reaction (2 ul, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Modification:

    Article Title: Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA
    Article Snippet: Cap analog and modified NTPs were purchased from Trilink Biotechnologies. .. To remove immunogenic 5′ triphosphate moieties from uncapped transcripts, 10 uL of Antarctic Phosphatase reaction buffer and 3 uL of Antarctic Phosphatase (NEB) was added to each prep.

    Western Blot:

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: IP efficiency was controlled with part of the sample by Western Blot as shown in . .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C.

    Electroporation:

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. 10 ng/μL cDNA plasmid template 10 μM For PCR primer 10 μM Rev PCR primer 10 μM For mutagenesis primer 10 μM Rev mutagenesis primer Herculase II Fusion DNA Polymerase 5× Herculase II reaction buffer (provided with Herculase II Fusion DNA Polymerase) dNTP mix (10 mM each dNTP) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Gel and PCR Cleanup Kit (e.g. Nucleospin Gel and PCR Clean-up, Machery-Nagel) ≥10 μg cDNA plasmid vector Restriction enzymes Buffer(s) for restriction enzymes (provided with enzyme(s)) BSA (if needed; provided with enzyme(s)) 5 U/μL Antarctic Phosphatase (NEB) 10× Antarctic Phosphatase Reaction Buffer (provided with 5U/μL Antarctic Phosphatase) Quick Ligation Kit (NEB; includes Quick T4 DNA Ligase and 2× Quick Ligation Reaction Buffer) SURE Electroporation-competent Cells (Agilent) Sterile water LB medium LB-Amp agar plates (50–100 μg/mL) LB-Amp medium (50–100 μg/mL) Mini-prep Kit (e.g. NucleoSpin Plasmid, Machery-Nagel) 1× TE (pH 8.0) Ice Electroporator and 1 mm cuvettes .. 1 Prepare 2 separate PCR reactions, one with each of these primer pairs: For PCR primer & Rev mutagenesis primer, For mutagenesis primer & Rev PCR primer.

    Chromatography:

    Article Title: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1
    Article Snippet: Paragraph title: Assay to determine dNTPase activity of SAMHD1 by thin-liquid chromatography ... The antarctic phosphatase reaction (2 ul, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Immunoprecipitation:

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: The cleared lysate was used for immunoprecipitation with rabbit IgG-conjugated Protein G magnetic beads (Invitrogen, Germany) on a rotating wheel for 4 hr or overnight at 4°C. .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C.

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: Immunoprecipitated and crosslinked RNA was partially digested with 50 U of RNase T1 per mL for 20 min at 25°C and 400 rpm. .. For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm.

    Generated:

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Substrate RNA oligonucleotides (below) that contained the suspected splice sites as shown by the RNAse H assays, were generated by run-off transcription and purified by 10% PAGE. .. Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs).

    Article Title: Improving the performance of true single molecule sequencing for ancient DNA
    Article Snippet: For each spiking experiment, we further assumed a binomial distribution with Rmin as the probability of success for estimating the number of expected endogenous sequences given the total number of sequences generated. .. For the phosphatase reactions, 0.5 μl of the DNA extracts were mixed with 8 μl of nuclease-free water, 1 μl of NEB Antarctic Phosphatase 10X Reaction buffer and 2.5 units of NEB Antarctic Phosphatase.

    other:

    Article Title: Predicting Silk Fiber Mechanical Properties through Multiscale Simulation and Protein Design
    Article Snippet: NEB endonuclease restriction enzyme buffer set (New England Biolabs, Ipswich, MA, USA) DNA Ligase Buffer (New England Biolabs, Ipswich, MA, USA) TAE (Tris-acetate-EDTA) buffer (Fisher Scientific, Pittsburgh, PA, USA) 10X Antarctic Phosphatase Reaction Buffer (New England Biolabs, Ipswich, MA, USA) NuPAGE MES SDS Running Buffer (20X) (ThermoFisher Scientific, Waltham, MA, USA) Denaturing buffer (8 M urea, 0.1 M NaH2PO4, 0.01 M Tris-HCl, pH 8.0)

    Polymerase Chain Reaction:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: The IVT reaction was performed at 37°C for 4 hr, and the mixture contained 7.5 mM ATP, 1.875 mM guanosine triphosphate (GTP) (both from the MEGAscript T7 kit), 7.5 mM 5-Methyl-CTP, 7.5 mM pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), 2.5 mM 3′-O-Me-m7G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany), 40 U RiboLock RNase inhibitor (Thermo Fisher Scientific), and 1.5 μg PCR product. .. Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. 10 ng/μL cDNA plasmid template 10 μM For PCR primer 10 μM Rev PCR primer 10 μM For mutagenesis primer 10 μM Rev mutagenesis primer Herculase II Fusion DNA Polymerase 5× Herculase II reaction buffer (provided with Herculase II Fusion DNA Polymerase) dNTP mix (10 mM each dNTP) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Gel and PCR Cleanup Kit (e.g. Nucleospin Gel and PCR Clean-up, Machery-Nagel) ≥10 μg cDNA plasmid vector Restriction enzymes Buffer(s) for restriction enzymes (provided with enzyme(s)) BSA (if needed; provided with enzyme(s)) 5 U/μL Antarctic Phosphatase (NEB) 10× Antarctic Phosphatase Reaction Buffer (provided with 5U/μL Antarctic Phosphatase) Quick Ligation Kit (NEB; includes Quick T4 DNA Ligase and 2× Quick Ligation Reaction Buffer) SURE Electroporation-competent Cells (Agilent) Sterile water LB medium LB-Amp agar plates (50–100 μg/mL) LB-Amp medium (50–100 μg/mL) Mini-prep Kit (e.g. NucleoSpin Plasmid, Machery-Nagel) 1× TE (pH 8.0) Ice Electroporator and 1 mm cuvettes .. 1 Prepare 2 separate PCR reactions, one with each of these primer pairs: For PCR primer & Rev mutagenesis primer, For mutagenesis primer & Rev PCR primer.

    Sonication:

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: Samples were solubilized for 1 min via sonication with a Covaris S220 instrument (Covaris, UK) using following parameters: Peak Incident Power (W): 140; Duty Factor: 5%; Cycles per Burst: 200. .. For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm.

    Recombinant:

    Article Title: Contribution of SAM and HD domains to retroviral restriction mediated by human SAMHD1
    Article Snippet: Recombinant SAMHD1 (5 µM) was incubated with or without 100 µM dGTP, with or without 6 µM dsRNA analog, 500 µM dTTP and 0.25 µl α32 P-dTTP (PerkinElmer) in SAMHD1 reaction buffer (50 mM Tris–HCl pH 8, 50 mM KCl, 5 mM MgCl2, 0.1% Triton-X 100) in a 17.5 µl final volume. .. The antarctic phosphatase reaction (2 µl, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Nucleic Acid Electrophoresis:

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C. .. Synthesized RNA products were then repurified using RNeasy Mini Columns (Qiagen) and quantitated by spectrophotometry and denaturing gel electrophoresis.

    Magnetic Beads:

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: The cleared lysate was used for immunoprecipitation with rabbit IgG-conjugated Protein G magnetic beads (Invitrogen, Germany) on a rotating wheel for 4 hr or overnight at 4°C. .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C.

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: Immunoprecipitation was performed on a rotating wheel overnight at 4°C with rabbit IgG-conjugated Protein G magnetic beads (Invitrogen, Germany). .. For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm.

    Mutagenesis:

    Article Title: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1
    Article Snippet: Assay to determine dNTPase activity of SAMHD1 by thin-liquid chromatography Wild type and mutant SAMHD1 proteins immunoprecipiated from mammalian cells were incubated with or without 100 μM dGTP, 500 μM dTTP and 0.25 μl α32P-dTTP (PerkinElmer) in SAMHD1 reaction buffer (50 mM Tris–HCl pH 8, 50 mM KCl, 5 mM MgCl2 , 0.1% Triton-X 100) in a 17.5 μl final volume. .. The antarctic phosphatase reaction (2 ul, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. 10 ng/μL cDNA plasmid template 10 μM For PCR primer 10 μM Rev PCR primer 10 μM For mutagenesis primer 10 μM Rev mutagenesis primer Herculase II Fusion DNA Polymerase 5× Herculase II reaction buffer (provided with Herculase II Fusion DNA Polymerase) dNTP mix (10 mM each dNTP) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Gel and PCR Cleanup Kit (e.g. Nucleospin Gel and PCR Clean-up, Machery-Nagel) ≥10 μg cDNA plasmid vector Restriction enzymes Buffer(s) for restriction enzymes (provided with enzyme(s)) BSA (if needed; provided with enzyme(s)) 5 U/μL Antarctic Phosphatase (NEB) 10× Antarctic Phosphatase Reaction Buffer (provided with 5U/μL Antarctic Phosphatase) Quick Ligation Kit (NEB; includes Quick T4 DNA Ligase and 2× Quick Ligation Reaction Buffer) SURE Electroporation-competent Cells (Agilent) Sterile water LB medium LB-Amp agar plates (50–100 μg/mL) LB-Amp medium (50–100 μg/mL) Mini-prep Kit (e.g. NucleoSpin Plasmid, Machery-Nagel) 1× TE (pH 8.0) Ice Electroporator and 1 mm cuvettes .. 1 Prepare 2 separate PCR reactions, one with each of these primer pairs: For PCR primer & Rev mutagenesis primer, For mutagenesis primer & Rev PCR primer.

    Labeling:

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs). .. Substrates were then 5′ radiolabeled at 37°C for 1 h. Twenty microliters labeling reactions consisted of 20 pmol RNA substrate, 20 μCi [γ-32 P]-ATP, 1× T4 Polynucleotide Kinase Reaction Buffer and 10 units T4 Polynucleotide Kinase (New England Biolabs).

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm. .. Phosphorylation of PAR-CLIP samples was performed using either 1 mM ATP per mL (cold-labeling) or 0.5 μCi of gamma-32-P-ATP per mL (radioactive labeling).

    Purification:

    Article Title: Aire controls gene expression in the thymic epithelium with ordered stochasticity
    Article Snippet: First the 5’ end of the aRNA was dephosphorylated by adding 4 µL of a mix containing 2µL of 10X Antarctic Phosphatase Reaction Buffer, 1µL of Antartic phosphatase (5U/µL stock, NEB M0289) and 1µL of RNAseOut to 16µL of each aRNA pool and incubating at 37°C for 30min and 65°C for 5min. .. The phosphatase and PNK treated RNA was then purified using RNeasy MinElute Cleanup Kit following the manufacturer’s protocol and eluted in 14µL of RNAse-free water.

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: Afterward, the template DNA was removed by adding 1 μL TURBO DNase (from the MEGAscript T7 kit) and incubation at 37°C for 15 min. Then the reaction mixture was purified using the RNeasy MinElute cleanup kit (QIAGEN) according to the manufacturer’s instructions. .. Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA
    Article Snippet: Reactions were incubated 4–6 hours at 37°C and treated with 2 uL TURBO DNase for a further 15 minutes at 37°C before being purified on MEGAclear (Ambion) spin columns, the RNA products being eluted in a volume of 100 uL. .. To remove immunogenic 5′ triphosphate moieties from uncapped transcripts, 10 uL of Antarctic Phosphatase reaction buffer and 3 uL of Antarctic Phosphatase (NEB) was added to each prep.

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Substrate RNA oligonucleotides (below) that contained the suspected splice sites as shown by the RNAse H assays, were generated by run-off transcription and purified by 10% PAGE. .. Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs).

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: Reactions were incubated 3-4 hours at 37°C then treated with 1 µl TURBO DNase (Life Technologies) for a further 15-minutes at 37°C before purification on RNeasy MiniSpin columns (Qiagen). .. To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C.

    Sequencing:

    Article Title: Improving the performance of true single molecule sequencing for ancient DNA
    Article Snippet: Paragraph title: tSMS sequencing ... For the phosphatase reactions, 0.5 μl of the DNA extracts were mixed with 8 μl of nuclease-free water, 1 μl of NEB Antarctic Phosphatase 10X Reaction buffer and 2.5 units of NEB Antarctic Phosphatase.

    De-Phosphorylation Assay:

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: .. Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs). .. Substrates were then 5′ radiolabeled at 37°C for 1 h. Twenty microliters labeling reactions consisted of 20 pmol RNA substrate, 20 μCi [γ-32 P]-ATP, 1× T4 Polynucleotide Kinase Reaction Buffer and 10 units T4 Polynucleotide Kinase (New England Biolabs).

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C. .. 3´ adapter ligation was performed in T4 RNA ligase buffer (NEB) with 10 U/µL T4 RNA ligase 2 (KQ) (NEB), 10 μM 3′ adapter (5′ 5rApp-TGGAATTCTCGGGTGCCAAGG-3ddC 3′ (IDT), 1 U/µL RNase OUT, and 15% (w/v) PEG 8000 overnight at 16°C.

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: .. For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm. ..

    Staining:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany). .. The quality and purity of the PCR product and the synthetic mRNA were assessed by 1% agarose gel electrophoresis and staining with 1× GelRed (Biotium, Fermont, USA).

    Concentration Assay:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany). .. The concentration of mRNA was determined using a photometer (BioPhotometer, Eppendorf, Hamburg, Germany).

    Article Title: Feeder-Free Derivation of Human Induced Pluripotent Stem Cells with Messenger RNA
    Article Snippet: To remove immunogenic 5′ triphosphate moieties from uncapped transcripts, 10 uL of Antarctic Phosphatase reaction buffer and 3 uL of Antarctic Phosphatase (NEB) was added to each prep. .. RNA yield was quantitated by Nanodrop (Thermo Scientific), and the preps were subsequently adjusted to a standardized working concentration of 100 ng/uL by addition of TE pH 7.0 (Ambion).

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C. .. 3´ adapter ligation was performed in T4 RNA ligase buffer (NEB) with 10 U/µL T4 RNA ligase 2 (KQ) (NEB), 10 μM 3′ adapter (5′ 5rApp-TGGAATTCTCGGGTGCCAAGG-3ddC 3′ (IDT), 1 U/µL RNase OUT, and 15% (w/v) PEG 8000 overnight at 16°C.

    Article Title: RNA-dependent chromatin association of transcription elongation factors and Pol II CTD kinases
    Article Snippet: For dephosphorylation, 1× antarctic phosphatase reaction buffer (NEB, Germany) with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) were added and the suspension was incubated at 37°C for 30 min and 800 rpm. .. Beads were resuspended in 1 × T4 PNK reaction buffer A (Fermentas, Germany) with a final concentration of 1 U/µL T4 PNK and 1 U/µL RNase OUT.

    Plasmid Preparation:

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. 10 ng/μL cDNA plasmid template 10 μM For PCR primer 10 μM Rev PCR primer 10 μM For mutagenesis primer 10 μM Rev mutagenesis primer Herculase II Fusion DNA Polymerase 5× Herculase II reaction buffer (provided with Herculase II Fusion DNA Polymerase) dNTP mix (10 mM each dNTP) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Gel and PCR Cleanup Kit (e.g. Nucleospin Gel and PCR Clean-up, Machery-Nagel) ≥10 μg cDNA plasmid vector Restriction enzymes Buffer(s) for restriction enzymes (provided with enzyme(s)) BSA (if needed; provided with enzyme(s)) 5 U/μL Antarctic Phosphatase (NEB) 10× Antarctic Phosphatase Reaction Buffer (provided with 5U/μL Antarctic Phosphatase) Quick Ligation Kit (NEB; includes Quick T4 DNA Ligase and 2× Quick Ligation Reaction Buffer) SURE Electroporation-competent Cells (Agilent) Sterile water LB medium LB-Amp agar plates (50–100 μg/mL) LB-Amp medium (50–100 μg/mL) Mini-prep Kit (e.g. NucleoSpin Plasmid, Machery-Nagel) 1× TE (pH 8.0) Ice Electroporator and 1 mm cuvettes .. 1 Prepare 2 separate PCR reactions, one with each of these primer pairs: For PCR primer & Rev mutagenesis primer, For mutagenesis primer & Rev PCR primer.

    Agarose Gel Electrophoresis:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany). .. The quality and purity of the PCR product and the synthetic mRNA were assessed by 1% agarose gel electrophoresis and staining with 1× GelRed (Biotium, Fermont, USA).

    In Vitro:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: The DNA product was then in vitro -transcribed into mRNA using the MEGAscript T7 kit (Thermo Fisher Scientific, Waltham, USA). .. Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Increased efficiency of evolved group I intron spliceozymes by decreased side product formation
    Article Snippet: Paragraph title: Identification of in vitro splicing side products with single-nucleotide resolution ( C) ... Substrates were dephosphorylated at 37°C for 1 h. Ten microliters of dephosphorylation reactions contained 2 ng substrate oligonucleotide, 1× Antarctic Phosphatase Reaction Buffer and 5 units Antarctic Phosphatase (New England Biolabs).

    Ethanol Precipitation:

    Article Title: Mapping 3D genome organization relative to nuclear compartments using TSA-Seq as a cytological ruler
    Article Snippet: .. Lamin B1 DamID DamID of lamin B1 was performed in K562 cells as described ( ; ) except that before the DpnI digestion, 1.5 µg genomic DNA was treated for 1 h with 5 U Antarctic phosphatase (M0289S; NEB) in 20 µl 1× Antarctic phosphatase reaction buffer (NEB) followed by ethanol precipitation. .. This additional step prevents ligation of the adaptor to DNA fragments originating from apoptotic cells.

    Ligation:

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C. .. 3´ adapter ligation was performed in T4 RNA ligase buffer (NEB) with 10 U/µL T4 RNA ligase 2 (KQ) (NEB), 10 μM 3′ adapter (5′ 5rApp-TGGAATTCTCGGGTGCCAAGG-3ddC 3′ (IDT), 1 U/µL RNase OUT, and 15% (w/v) PEG 8000 overnight at 16°C.

    Article Title: Poliovirus: Generation and Characterization of Mutants
    Article Snippet: .. 10 ng/μL cDNA plasmid template 10 μM For PCR primer 10 μM Rev PCR primer 10 μM For mutagenesis primer 10 μM Rev mutagenesis primer Herculase II Fusion DNA Polymerase 5× Herculase II reaction buffer (provided with Herculase II Fusion DNA Polymerase) dNTP mix (10 mM each dNTP) Nuclease-free water 1% agarose TAE gels and running buffer ( ) 6× DNA Loading Dye DNA Ladder (e.g. GeneRuler 1 kb Plus, Fermentas) Gel and PCR Cleanup Kit (e.g. Nucleospin Gel and PCR Clean-up, Machery-Nagel) ≥10 μg cDNA plasmid vector Restriction enzymes Buffer(s) for restriction enzymes (provided with enzyme(s)) BSA (if needed; provided with enzyme(s)) 5 U/μL Antarctic Phosphatase (NEB) 10× Antarctic Phosphatase Reaction Buffer (provided with 5U/μL Antarctic Phosphatase) Quick Ligation Kit (NEB; includes Quick T4 DNA Ligase and 2× Quick Ligation Reaction Buffer) SURE Electroporation-competent Cells (Agilent) Sterile water LB medium LB-Amp agar plates (50–100 μg/mL) LB-Amp medium (50–100 μg/mL) Mini-prep Kit (e.g. NucleoSpin Plasmid, Machery-Nagel) 1× TE (pH 8.0) Ice Electroporator and 1 mm cuvettes .. 1 Prepare 2 separate PCR reactions, one with each of these primer pairs: For PCR primer & Rev mutagenesis primer, For mutagenesis primer & Rev PCR primer.

    Article Title: Mapping 3D genome organization relative to nuclear compartments using TSA-Seq as a cytological ruler
    Article Snippet: Lamin B1 DamID DamID of lamin B1 was performed in K562 cells as described ( ; ) except that before the DpnI digestion, 1.5 µg genomic DNA was treated for 1 h with 5 U Antarctic phosphatase (M0289S; NEB) in 20 µl 1× Antarctic phosphatase reaction buffer (NEB) followed by ethanol precipitation. .. This additional step prevents ligation of the adaptor to DNA fragments originating from apoptotic cells.

    Spectrophotometry:

    Article Title: mRNA Transfection of Mouse and Human Neural Stem Cell Cultures
    Article Snippet: To remove 5′ triphosphate moieties from uncapped transcripts, 10 µl of Antarctic Phosphatase reaction buffer and 5 µl (25U) of Antarctic Phosphatase (New England BioLabs) was added to each RNA product and incubated for 30-minutes at 37°C. .. Synthesized RNA products were then repurified using RNeasy Mini Columns (Qiagen) and quantitated by spectrophotometry and denaturing gel electrophoresis.

    Activation Assay:

    Article Title: Intradermal Delivery of Synthetic mRNA Using Hollow Microneedles for Efficient and Rapid Production of Exogenous Proteins in Skin
    Article Snippet: PCR was run using the following cycling protocol: initial activation step at 94°C for 3 min, followed by 25 cycles of denaturation at 94°C for 45 s, annealing at 55°C for 1 min, extension at 72°C for 1 min, and final extension at 72°C for 5 min. During the PCR, a poly T-tail of 120 thymidines (T) was added to the insert. .. Afterward, the mRNA was treated for 30 min at 37°C with 5 U Antarctic phosphatase and 1× Antarctic phosphatase reaction buffer in a total volume of 45.5 μl (New England Biolabs, Frankfurt am Main, Germany).

    Mobility Shift:

    Article Title: Phosphorylation of ASC acts as a molecular switch controlling the formation of speck-like aggregates and inflammasome activity
    Article Snippet: Paragraph title: Phos-tag-based Mobility Shift Assay ... Proteins in the cell lysates were precipitated with acetone, incubated in phosphatase reaction mixture containing 250 U/ml of antarctic phosphatase (New England Biolabs) overnight at 37°C, reprecipitated with acetone and dissolved in SDS sample buffer.

    Thin Layer Chromatography:

    Article Title: Contribution of SAM and HD domains to retroviral restriction mediated by human SAMHD1
    Article Snippet: The antarctic phosphatase reaction (2 µl, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility. .. Reactions were spotted (0.5 µl) on a TLC PEI Cellulose F plate (EMD Chemicals) and separated in a 0.8 M LiCl solvent.

    Article Title: Contribution of oligomerization to the anti-HIV-1 properties of SAMHD1
    Article Snippet: The antarctic phosphatase reaction (2 ul, New England BioLabs) was used to show the mobility of monophosphates on the plate as a comparison to triphosphate mobility. .. Reactions were spotted (0.5 μl) on a TLC PEI Cellulose F plate (EMD Chemicals) and separated in a 0.8 M LiCl solvent.

    Lysis:

    Article Title: Transcriptome maps of general eukaryotic RNA degradation factors
    Article Snippet: Lysis was performed in lysis buffer (50 mM Tris-HCl pH 7.5, 100 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS, 0.5% NP-40) by bead beating (FastPrep−24 Instrument, MP Biomedicals, LLC., France) using silica-zirconium beads (Roth, Germany). .. The dephosphorylation reaction was performed in antarctic phosphatase reaction buffer (NEB, Germany) supplemented with 1 U/µL of antarctic phosphatase and 1 U/µL of RNase OUT (Invitrogen) at 37°C for 30 min. For rephosphyorylation, beads were incubated in T4 PNK reaction buffer A (Invitrogen) with a final concentration of 1 U/µL T4 PNK, 1 U/µL RNase OUT and 1 mM ATP for 1 hr at 37°C.

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    New England Biolabs antarctic phosphatase reaction buffer
    Antarctic Phosphatase Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antarctic phosphatase reaction buffer/product/New England Biolabs
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    antarctic phosphatase reaction buffer - by Bioz Stars, 2020-01
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