presence of biotin 11 dctp  (Jena Bioscience)


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    Name:
    Biotin 11 dCTP
    Description:

    Catalog Number:
    NU-809-BIOX-L
    Price:
    239.94
    Category:
    Nucleotides Nucleosides
    Size:
    5 x 200 µl
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    Structured Review

    Jena Bioscience presence of biotin 11 dctp
    Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of <t>Biotin-11-dCTP</t> for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.

    https://www.bioz.com/result/presence of biotin 11 dctp/product/Jena Bioscience
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    presence of biotin 11 dctp - by Bioz Stars, 2021-06
    96/100 stars

    Images

    1) Product Images from "The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication"

    Article Title: The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication

    Journal: bioRxiv

    doi: 10.1101/2021.03.12.435139

    Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.
    Figure Legend Snippet: Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.

    Techniques Used: Activity Assay, In Vitro, Labeling, Incubation, Recombinant, DNA Synthesis

    2) Product Images from "The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication"

    Article Title: The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication

    Journal: bioRxiv

    doi: 10.1101/2021.03.12.435139

    Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.
    Figure Legend Snippet: Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.

    Techniques Used: Activity Assay, In Vitro, Labeling, Incubation, Recombinant, DNA Synthesis

    Related Articles

    Incubation:

    Article Title: APOBEC3G Inhibits Elongation of HIV-1 Reverse Transcripts
    Article Snippet: For later experiments, an equal volume of 2× Cells-to-signal lysis buffer (Ambion) was added instead of PBS with carrier DNA, and samples were diluted 1∶10 in water before analysing 2 µl directly in real-time quantitative PCR. .. Reverse Transcription Initiation nERT Assay For this assay, a nERT reaction was performed essentially as above, with the exception of the dNTP addition: pelletted virions were resuspended in PBS, 2.5 mM MgCl2 , 15 µg/ml melittin and 0.5mM biotin-11-dCTP (Jena Bioscience), and incubated at 37°C. .. Control reactions were incubated without any dNTPs for 2 h. At the specified time points, aliquots were removed, added to QIAzol lysis buffer (Qiagen) and frozen at −80°C.

    Activity Assay:

    Article Title: Cohesin residency determines chromatin loop patterns
    Article Snippet: Tight binding and chemical crosslinking between DNA and nucleosome are supposed to prevent the Klenow enzyme’s 3’-to-5’ exonuclease activity from chewing nucleosomal DNA all the way through. .. Third, Klenow enzyme’s polymerase activity repairs the nucleosomal DNA ends to the blunt and ligatible ends upon the supplement of dNTPs including 66uM dTTP, dGTP (NEB #N0446), biotin-dATP (Jena Bioscience #NU-835-BIO14), and biotin-dCTP (Jena Bioscience #NU-809-BIOX) in 1X T4 DNA ligase reaction buffer (NEB #B0202). .. The reaction was incubated at 25°C for 45 minutes with interval mixing and then inactivated with 30mM EDTA at 65°C for 20 minutes.

    Concentration Assay:

    Article Title: Atomic-resolution mapping of transcription factor-DNA interactions by femtosecond laser crosslinking and mass spectrometry
    Article Snippet: Forty-five microliters of T4 PNK (New England Biolabs, M0201S) and five microliters NEBuffer 2 (+100 µg ml−1 BSA) were added to 45 µg of cross-linked or non-cross-linked chromatin, respectively. .. For the biotin-replacement synthesis, the following reagents were added to 45 µg of cross-linked or 45 µg non-cross-linked chromatin at a concentration of 0.5 µg µl−1 , respectively: 3.1 µl of 10 mg ml−1 BSA (New England Biolabs), 21 µl of 10× NEB buffer 2 (New England Biolabs), 76.3 µl of 0.4 mM Biotin-dATP (Jena Biosciences), 76.3 µl of 0.4 mM Biotin-dCTP (Jena Biosciences), 3.1 µl of 10 mM dTTP/dGTP (New England Biolabs), and 30 µl of T4 Polymerase at a concentration of 3 U µl−1 (New England Biolabs, M0203S). .. After incubation at 12 °C for 15 min, EDTA was added to a final concentration of 50 mM.

    Article Title: Resolving the 3D landscape of transcription-linked mammalian chromatin folding
    Article Snippet: .. The blunting and labeling reaction was triggered upon adding biotin-dATP (Jena Bioscience # NU-835-BIO14), biotin-dCTP (Jena Bioscience # NU-809-BIOX), dGTP, and dTTP to a final concentration of 66 mM each. .. Incubation for 45 minutes at room temperature is sufficient to convert most MNase-digested ends to blunt ends for proximity ligation.

    In Vitro:

    Article Title: The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication
    Article Snippet: The relative level of Biotin-16-UTP in each reaction was calculated and plotted. .. In vitro RNA-primed DNA replication The in vitro RNA-primed DNA replication assay was carried out using M13mp18 ssDNA as the template and in the presence of Biotin-11-dCTP (Jena Bioscience). .. Reactions (20 µl) contained 35 fmol M13mp18 ssDNA, 10.5 mM MgCl2 , 25 mM Tris-HCl (pH 8.0), 1 mM DTT, 100 µg/ml BSA, 400 µM ATP, 150 µM GTP, UTP and CTP, 100 µM dATP, dGTP and dTTP, 60 µM dCTP, and 40 µM Biotin-11-dCTP, 300 fmol POL γ and 500 fmol POLRMT or 500 fmol POLRMTE423P.

    Labeling:

    Article Title: Resolving the 3D landscape of transcription-linked mammalian chromatin folding
    Article Snippet: .. The blunting and labeling reaction was triggered upon adding biotin-dATP (Jena Bioscience # NU-835-BIO14), biotin-dCTP (Jena Bioscience # NU-809-BIOX), dGTP, and dTTP to a final concentration of 66 mM each. .. Incubation for 45 minutes at room temperature is sufficient to convert most MNase-digested ends to blunt ends for proximity ligation.

    other:

    Article Title: The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication
    Article Snippet: Reactions (20 µl) contained 35 fmol M13mp18 ssDNA, 10.5 mM MgCl2 , 25 mM Tris-HCl (pH 8.0), 1 mM DTT, 100 µg/ml BSA, 400 µM ATP, 150 µM GTP, UTP and CTP, 100 µM dATP, dGTP and dTTP, 60 µM dCTP, and 40 µM Biotin-11-dCTP, 300 fmol POL γ and 500 fmol POLRMT or 500 fmol POLRMTE423P.

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  • 96
    Jena Bioscience presence of biotin 11 dctp
    Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of <t>Biotin-11-dCTP</t> for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.
    Presence Of Biotin 11 Dctp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/presence of biotin 11 dctp/product/Jena Bioscience
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    presence of biotin 11 dctp - by Bioz Stars, 2021-06
    96/100 stars
      Buy from Supplier

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    Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.

    Journal: bioRxiv

    Article Title: The PPR domain of mitochondrial RNA polymerase is a ribonuclease required for mtDNA replication

    doi: 10.1101/2021.03.12.435139

    Figure Lengend Snippet: Exoribonuclease activity of POLRMT is essential for priming DNA replication in vitro . (A) POLRMT E423P lacks the exoribonuclease activity. 5’-Biotin-labeled target ssRNA (0.05 µM) was incubated with recombinant POLRMT or POLRMT E423P (0.5 µM) in 10 µl reaction mix for indicated time (min) and separated on a denaturing polyacrylamide gel (20%). Arrowhead indicates the full-length target RNA. (B) Pseudo-first-order cleavage kinetics of 5’-Biotin-labeled target ssRNA by POLRMT and POLRMT E423P . The level of remaining RNA in each time point was measured, normalized to the initial level and plotted. Each data point represents the average of three independent experiments. Error bars represent sd. (C) RNA synthesis of POLRMT, POLRMT E423P (500 fmol) and T7 RNA polymerase (0.5 units) on M13mp18 ssDNA template. RNA products were separated on a denaturing polyacrylamide gel (10%). Molecular size markers are shown on the left. (D) Kinetics of Biotin-16-UTP incorporation in the in vitro transcription assay using POLRMT, POLRMT E423P (500 fmol) or T7 RNA polymerase (0.5 units) on ssDNA template. Each time point represents the average of three independent experiments. Error bars represent sd. A line of linear regression was plotted for each protein. (POLRMT, R 2 =0.98; POLRMT E423P , R 2 =0.84; T7 RNA polymerase, R 2 =0.96.) (E) The RNA-primed DNA synthesis assay was monitored in the presence of Biotin-11-dCTP for labeling of DNA products. Lane 1, POLRMT (500 fmol) and POL γ (300 fmol); Lane 2, POLRMT E423P (500 fmol) and POL γ (300 fmol); Lane 3, Control experiment using POL γ (300 fmol) only. Reactions were incubated for 1 h at 37 °C and the products were analyzed on a denaturing polyacrylamide gel (10%). Each data point represents the average of three independent experiments. Error bars represent sd.

    Article Snippet: In vitro RNA-primed DNA replication The in vitro RNA-primed DNA replication assay was carried out using M13mp18 ssDNA as the template and in the presence of Biotin-11-dCTP (Jena Bioscience).

    Techniques: Activity Assay, In Vitro, Labeling, Incubation, Recombinant, DNA Synthesis