phi29 dna polymerase  (Thermo Fisher)


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    Structured Review

    Thermo Fisher phi29 dna polymerase
    Loss of SLX4IP in ALT-Positive Cells Increases ALT-Related Phenotypes (A) Genomic <t>DNA</t> was isolated from U2OS cells and processed to detect <t>Phi29-dependent</t> telomere circles. The Phi29 amplification products were detected by Southern blotting using a γ[ 32 P]-labeled telomeric (TTAGGG) probe. (B) Quantification of (A). The extent of [ 32 P] incorporation was quantified from the autoradiograph and normalized to SLX4IP +/+ , which was arbitrarily assigned a value of 1. Data are represented as mean ± SD; n = 3; ∗ p
    Phi29 Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phi29 dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    phi29 dna polymerase - by Bioz Stars, 2020-01
    99/100 stars

    Images

    1) Product Images from "SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance"

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance

    Journal: Molecular Cell

    doi: 10.1016/j.molcel.2019.07.010

    Loss of SLX4IP in ALT-Positive Cells Increases ALT-Related Phenotypes (A) Genomic DNA was isolated from U2OS cells and processed to detect Phi29-dependent telomere circles. The Phi29 amplification products were detected by Southern blotting using a γ[ 32 P]-labeled telomeric (TTAGGG) probe. (B) Quantification of (A). The extent of [ 32 P] incorporation was quantified from the autoradiograph and normalized to SLX4IP +/+ , which was arbitrarily assigned a value of 1. Data are represented as mean ± SD; n = 3; ∗ p
    Figure Legend Snippet: Loss of SLX4IP in ALT-Positive Cells Increases ALT-Related Phenotypes (A) Genomic DNA was isolated from U2OS cells and processed to detect Phi29-dependent telomere circles. The Phi29 amplification products were detected by Southern blotting using a γ[ 32 P]-labeled telomeric (TTAGGG) probe. (B) Quantification of (A). The extent of [ 32 P] incorporation was quantified from the autoradiograph and normalized to SLX4IP +/+ , which was arbitrarily assigned a value of 1. Data are represented as mean ± SD; n = 3; ∗ p

    Techniques Used: Isolation, Amplification, Southern Blot, Labeling, Autoradiography

    SLX4 Depletion Further Augments the Increase in ALT-Related Phenotypes in SLX4IP −/− Cells (A) U2OS cells were transfected with the indicated siRNAs. Their genomic DNA was then processed to detect Phi29-dependent telomere circles. The Phi29 amplification products were detected by Southern blotting using a γ[ 32 P]-labeled telomeric (TTAGGG) probe. (B) Quantification of (A). The extent of [ 32 P] incorporation was quantified from the autoradiograph and normalized to SLX4IP +/+ siCTRL, which was arbitrarily assigned a value of 1. Data are represented as mean ± SD; n = 3; ∗ p
    Figure Legend Snippet: SLX4 Depletion Further Augments the Increase in ALT-Related Phenotypes in SLX4IP −/− Cells (A) U2OS cells were transfected with the indicated siRNAs. Their genomic DNA was then processed to detect Phi29-dependent telomere circles. The Phi29 amplification products were detected by Southern blotting using a γ[ 32 P]-labeled telomeric (TTAGGG) probe. (B) Quantification of (A). The extent of [ 32 P] incorporation was quantified from the autoradiograph and normalized to SLX4IP +/+ siCTRL, which was arbitrarily assigned a value of 1. Data are represented as mean ± SD; n = 3; ∗ p

    Techniques Used: Transfection, Amplification, Southern Blot, Labeling, Autoradiography

    Related Articles

    Nucleic Acid Electrophoresis:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Primer extension was carried out with 7.5 U of Phi29 DNA polymerase (Thermo Scientific) at 30°C for 12 hours. .. The extension products were separated by denaturing gel electrophoresis (0.8% agarose, 50 mM NaOH, and 1 mM EDTA [pH 8]) at 2 V/cm for 18 hours and transferred onto a nylon membrane (GE Healthcare) in 10X SSC.

    Incubation:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: T-circle assay To isolate genomic DNA, cells were then resuspended in TNE (10 mM Tris pH 7.4, 10 mM EDTA, 100 mM NaCl) and lysed in TNES (10 mM Tris pH 7.4, 100 mM NaCl, 10 mM EDTA, 1% SDS) in the presence of 100 μg/ml proteinase K. After overnight incubation with proteinase K at 37°C, and phenol/chloroform extractions, DNA was precipitated with isopropanol and resuspended in TE (10 mM Tris pH 7.5/1 mM EDTA). .. Primer extension was carried out with 7.5 U of Phi29 DNA polymerase (Thermo Scientific) at 30°C for 12 hours.

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    Thermo Fisher phi29 dna polymerase
    Size distribution of <t>DNA-magnesium-pyrophosphate</t> particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate <t>phi29</t> DNA polymerase and terminate the MDA reaction.
    Phi29 Dna Polymerase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phi29 dna polymerase/product/Thermo Fisher
    Average 90 stars, based on 50 article reviews
    Price from $9.99 to $1999.99
    phi29 dna polymerase - by Bioz Stars, 2020-01
    90/100 stars
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    Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Journal: Micromachines

    Article Title: Droplet Microfluidics Approach for Single-DNA Molecule Amplification and Condensation into DNA-Magnesium-Pyrophosphate Particles

    doi: 10.3390/mi8020062

    Figure Lengend Snippet: Size distribution of DNA-magnesium-pyrophosphate particles as a function of multiple displacement amplification reaction times, with and without the heat-inactivation step. At different time points (17, 38 and 66 h) of the MDA reaction in 3-pL droplets at 30 °C, the resulting DNA-Mg-PP i particles were released from droplets and imaged under TEM to measure their size. The first set of measurements (green) was made without heat-inactivation step. The second set of measurements (grey) was made with heat-inactivation step at 65 °C for 15 min that is typically used to inactivate phi29 DNA polymerase and terminate the MDA reaction.

    Article Snippet: Single-DNA Molecule Encapsulation and Amplification The MDA reaction mix contained pIVEX2.2-lacZ-his plasmid, 1× phi29 reaction buffer (33 mM Tris-acetate (pH 7.9), 10 mM Mg-acetate, 66 mM K-acetate, 0.1% (v /v ) Tween 20, 1 mM dithiothreitol (DTT)), 50 μM exo-nuclease resistant hexanucleotide primers, 1 mM of each deoxynucleoside triphosphates (dNTP), 0.4% (w /v ) Pluronic F-127 and 0.8 U/μL phi29 DNA polymerase (Thermo Fisher Scientific, Vilnius, Lithuania).

    Techniques: Multiple Displacement Amplification, Transmission Electron Microscopy