phi29 dna polymerase 10 u µl  (Thermo Fisher)


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    Thermo Fisher phi29 dna polymerase 10 u µl

    https://www.bioz.com/result/phi29 dna polymerase 10 u µl/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    phi29 dna polymerase 10 u µl - by Bioz Stars, 2020-09
    99/100 stars

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    Article Title: Manipulating the hydrophobicity of DNA as a universal strategy for visual biosensing.
    Article Snippet: Current visual biosensing methods, including colorimetric-based, fluorescence-based and chemiluminescence-based methods, are inappropriate for the hundreds of millions of people affected by color blindness and color weakness.

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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: 99/100, based on 62 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 62 article reviews
    Price from $9.99 to $1999.99
    phi29 dna polymerase - by Bioz Stars, 2020-09
    99/100 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: In this context, DNA amplification driven by phi29 DNA polymerase provides an alternative approach to amplify long DNA molecules and because of isothermal reaction conditions [ , ], the potential problems associated with emulsion stability become irrelevant.

    Techniques: Multiple Displacement Amplification, Transmission Electron Microscopy

    Real-time reverse transcription–template dependent multiple displacement amplification (RT-tdMDA) using three hepatitis C virus (HCV) patient serum samples and a negative control (H 2 O) These samples covered the range of the RNA yield extracted from 200 μL serum (7.5–19.2 ng), as quantitated in the final 14 μL of elution by the QIAGEN miRNA kit prior to HL-DNase digestion. An aliquot of 10.6 μL RNA was used for RT in a reaction containing 200 U SuperScript III, 80 μM 5′-end-blocked random pentamer primer (5′-/iSpC3/NNN*N*N-3′; asterisks denote phosphorothioate bonds), and 2 mM dNTPs in a 20-μL volume. An aliquot of 4 μL of the RT reaction was used in a 40-μL tdMDA reaction containing 300 U phi29 DNA polymerase (Epicentre), 80 μM primer, and 0.1× SYBR Green I (Thermo Fisher Scientific). The reaction was incubated at 28°C for 24 h on the ABI TaqMan 7500, in which fluorescent intensities were monitored through the SYBR Green channel. Estimated amount of cDNA input in tdMDA = [10.6 / (14 + 0.5 (HL-DNase) + 1.4 (buffer)] × [total RNA amount] × 0.2. Template-independent amplification was completely inhibited, as indicated by the negative control.

    Journal: BioTechniques

    Article Title: Template-dependent multiple displacement amplification for profiling human circulating RNA

    doi: 10.2144/000114566

    Figure Lengend Snippet: Real-time reverse transcription–template dependent multiple displacement amplification (RT-tdMDA) using three hepatitis C virus (HCV) patient serum samples and a negative control (H 2 O) These samples covered the range of the RNA yield extracted from 200 μL serum (7.5–19.2 ng), as quantitated in the final 14 μL of elution by the QIAGEN miRNA kit prior to HL-DNase digestion. An aliquot of 10.6 μL RNA was used for RT in a reaction containing 200 U SuperScript III, 80 μM 5′-end-blocked random pentamer primer (5′-/iSpC3/NNN*N*N-3′; asterisks denote phosphorothioate bonds), and 2 mM dNTPs in a 20-μL volume. An aliquot of 4 μL of the RT reaction was used in a 40-μL tdMDA reaction containing 300 U phi29 DNA polymerase (Epicentre), 80 μM primer, and 0.1× SYBR Green I (Thermo Fisher Scientific). The reaction was incubated at 28°C for 24 h on the ABI TaqMan 7500, in which fluorescent intensities were monitored through the SYBR Green channel. Estimated amount of cDNA input in tdMDA = [10.6 / (14 + 0.5 (HL-DNase) + 1.4 (buffer)] × [total RNA amount] × 0.2. Template-independent amplification was completely inhibited, as indicated by the negative control.

    Article Snippet: First, using the standard MDA protocol, we evaluated the purity of phi29 DNA polymerases from various vendors, including Lucigen (Middleton, WI), Epicentre (Madison, WI), Thermo Scientific (St. Louis, MO), and New England Biolabs (Ipswich, MA).

    Techniques: Multiple Displacement Amplification, Negative Control, SYBR Green Assay, Incubation, Amplification

    Effect of TEN1 depletion on T-circle release. A and B , Representative gels showing Phi29 amplification products obtained with DNA from shTEN1 or shSTN1 HeLa 1.2.11 ( A ) or shTEN1 HCT116 ( B ) cells relative to shNT, shTEN1 rescue, or U2OS controls. The amounts

    Journal: The Journal of Biological Chemistry

    Article Title: Human TEN1 Maintains Telomere Integrity and Functions in Genome-wide Replication Restart *

    doi: 10.1074/jbc.M113.493478

    Figure Lengend Snippet: Effect of TEN1 depletion on T-circle release. A and B , Representative gels showing Phi29 amplification products obtained with DNA from shTEN1 or shSTN1 HeLa 1.2.11 ( A ) or shTEN1 HCT116 ( B ) cells relative to shNT, shTEN1 rescue, or U2OS controls. The amounts

    Article Snippet: 0.2 m m dNTP was added in the presence or absence of Phi29 polymerase (Thermo) and incubated at 30 °C for 12 h followed by 65 °C for 20 min. Products were separated in 0.6% agarose gels at 100V for 1 h followed by 35 V overnight.

    Techniques: Amplification

    Schematic illustration of in situ PLA using conventional and UnFold probes. ( a ) Conventional in situ PLA. ( b ) In situ PLA using UnFold probes. (i) After pairs of primary antibodies have bound a pair of interacting proteins (red and green) followed by washes, secondary conventional or UnFold in situ PLA probes are added, followed after an incubation by renewed washes. (ii) In the conventional design under ( a ) two more oligonucleotides are then added that can form a DNA circle. Using the UnFold design in ( b ) the probe carrying a hairpin-loop oligonucleotide is cleaved at the U residues, liberating a free 5′ end capable of being ligated to the 3′ end of the same DNA strand. Meanwhile, the U residues in the hairpin DNA strand of the other UnFold probe are cleaved presenting a single-stranded template for the enzymatic joining of the ends of the strand on the first UnFold probe. (iii) A DNA ligase is added to form DNA circles in the two variants of in situ PLA. (iv) Finally, phi29 DNA polymerase is added to initiate RCA primed by oligonucleotides on one of the antibodies, and fluorescent oligonucleotides are used to visualize the RCA products.

    Journal: Scientific Reports

    Article Title: Improved efficiency of in situ protein analysis by proximity ligation using UnFold probes

    doi: 10.1038/s41598-018-23582-1

    Figure Lengend Snippet: Schematic illustration of in situ PLA using conventional and UnFold probes. ( a ) Conventional in situ PLA. ( b ) In situ PLA using UnFold probes. (i) After pairs of primary antibodies have bound a pair of interacting proteins (red and green) followed by washes, secondary conventional or UnFold in situ PLA probes are added, followed after an incubation by renewed washes. (ii) In the conventional design under ( a ) two more oligonucleotides are then added that can form a DNA circle. Using the UnFold design in ( b ) the probe carrying a hairpin-loop oligonucleotide is cleaved at the U residues, liberating a free 5′ end capable of being ligated to the 3′ end of the same DNA strand. Meanwhile, the U residues in the hairpin DNA strand of the other UnFold probe are cleaved presenting a single-stranded template for the enzymatic joining of the ends of the strand on the first UnFold probe. (iii) A DNA ligase is added to form DNA circles in the two variants of in situ PLA. (iv) Finally, phi29 DNA polymerase is added to initiate RCA primed by oligonucleotides on one of the antibodies, and fluorescent oligonucleotides are used to visualize the RCA products.

    Article Snippet: The slides were then washed twice in TBS-T for 3 min. Circularization oligonucleotides for the two probe designs were ligated using 0.02 U/µl T4 DNA ligase (Thermo Scientific) in T4 DNA ligase buffer supplemented with 0.25 mg/ml BSA for 30 min at 37 °C, thereafter the slides were washed twice for 3 min in TBS-T. RCA was performed by addition of 0.5 U/µl phi29 polymerase (Thermo scientific) in phi29 polymerase buffer (Thermo Scientific) supplemented with 7.5 ng/ml PolyA (Sigma-Aldrich), 0.25 mM dNTP and 0.25 mg/ml BSA for 60 min at 37 °C, followed by two 3 min washes in TBS-T.

    Techniques: In Situ, Proximity Ligation Assay, Incubation