platinium taq dna polymerase  (Thermo Fisher)


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    Name:
    Platinum Taq DNA Polymerase
    Description:
    Invitrogen Platinum Taq DNA Polymerase is a convenient and reliable hot start thermostable DNA polymerase for PCR that provides enhanced specificity over that of Taq DNA Polymerase The hot start property of the enzyme is conferred by thermolabile monoclonal antibodies that render Taq DNA polymerase inactive until the initial PCR denaturation step thus preventing the extention of nonspecifically annealed primers and improving product yield Using Platinum Taq DNA Polymerase The hot start property of Platinum Taq DNA Polymerase allows for convenient reaction assembly at room temperature Just as with Taq DNA Polymerase Platinum Taq DNA Polymerase has a non template dependent terminal transferase activity that adds a 3 deoxyadenosine to product ends and has a 5 →3 exonuclease activity PCR products generated with Platinum Taq DNA Polymerase may be used in the same downstream applications without protocol modifications Use Platinum Taq DNA Polymerase for the amplification of DNA from complex genomic viral and plasmid templates as well as in RT PCR Note For superior PCR performance we recommend next generation enzyme Platinum II Taq Hot start DNA Polymerase Platinum II Taq Hot Start DNA Polymerase is designed for universal primer annealing and fast easy PCR with its unique combination of innovative buffer high performance engineered Taq DNA polymerase and superior hot start technology
    Catalog Number:
    10966018
    Price:
    None
    Applications:
    Amplification of Bisulfite-Treated DNA|Chromatin Immunoprecipitation (ChIP)|Fast PCR|Hot Start PCR|PCR|PCR & Real-Time PCR|PCR Enzymes & Master Mixes|RNAi, Epigenetics & Non-Coding RNA Research|Routine PCR|Chromatin Biology|Methylation Analysis
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher platinium taq dna polymerase
    MNN2 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 23 colonies for each condition. Dark grey: Petri dishes cultures, light grey: liquid cultures. B ) Average of the 3 assays presented in A (standard deviation=19.55). C ) PCR product visualized on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications have been carried out with the <t>Platinium</t> <t>Taq.</t>
    Invitrogen Platinum Taq DNA Polymerase is a convenient and reliable hot start thermostable DNA polymerase for PCR that provides enhanced specificity over that of Taq DNA Polymerase The hot start property of the enzyme is conferred by thermolabile monoclonal antibodies that render Taq DNA polymerase inactive until the initial PCR denaturation step thus preventing the extention of nonspecifically annealed primers and improving product yield Using Platinum Taq DNA Polymerase The hot start property of Platinum Taq DNA Polymerase allows for convenient reaction assembly at room temperature Just as with Taq DNA Polymerase Platinum Taq DNA Polymerase has a non template dependent terminal transferase activity that adds a 3 deoxyadenosine to product ends and has a 5 →3 exonuclease activity PCR products generated with Platinum Taq DNA Polymerase may be used in the same downstream applications without protocol modifications Use Platinum Taq DNA Polymerase for the amplification of DNA from complex genomic viral and plasmid templates as well as in RT PCR Note For superior PCR performance we recommend next generation enzyme Platinum II Taq Hot start DNA Polymerase Platinum II Taq Hot Start DNA Polymerase is designed for universal primer annealing and fast easy PCR with its unique combination of innovative buffer high performance engineered Taq DNA polymerase and superior hot start technology
    https://www.bioz.com/result/platinium taq dna polymerase/product/Thermo Fisher
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    platinium taq dna polymerase - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae"

    Article Title: PCR on yeast colonies: an improved method for glyco-engineered Saccharomyces cerevisiae

    Journal: BMC Research Notes

    doi: 10.1186/1756-0500-6-201

    MNN2 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 23 colonies for each condition. Dark grey: Petri dishes cultures, light grey: liquid cultures. B ) Average of the 3 assays presented in A (standard deviation=19.55). C ) PCR product visualized on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications have been carried out with the Platinium Taq.
    Figure Legend Snippet: MNN2 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 23 colonies for each condition. Dark grey: Petri dishes cultures, light grey: liquid cultures. B ) Average of the 3 assays presented in A (standard deviation=19.55). C ) PCR product visualized on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications have been carried out with the Platinium Taq.

    Techniques Used: Amplification, Modification, Standard Deviation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    Efficiency of MNN5 gene amplification from wild type and modified strains. A ) Percentage of colonies picked from Petri dishes that amplified MNN5. The experiment was carried out on 47 colonies for each strain. Dark grey: BY4742; light grey: YiMMOgène. Amplification was carried out using the DreamTaq or the Platinium Taq. B ). PCR product visualisation, after amplification with the Platinium Taq, on a 1% agarose gel stained with SYBR safe. Left panel (1): BY4742, right panel (2): YiMMOgène, ML: Molecular ladder.
    Figure Legend Snippet: Efficiency of MNN5 gene amplification from wild type and modified strains. A ) Percentage of colonies picked from Petri dishes that amplified MNN5. The experiment was carried out on 47 colonies for each strain. Dark grey: BY4742; light grey: YiMMOgène. Amplification was carried out using the DreamTaq or the Platinium Taq. B ). PCR product visualisation, after amplification with the Platinium Taq, on a 1% agarose gel stained with SYBR safe. Left panel (1): BY4742, right panel (2): YiMMOgène, ML: Molecular ladder.

    Techniques Used: Amplification, Modification, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    Efficiency of MNN5 gene amplification from liquid culture of modified strain. A ) Percentage of MNN5 amplification from 47 colonies picked on Petri dishes and 47 colonies picked from liquid cultures in micro-plates. B ) Growth curve in micro-plate (YPD medium), average of two colonies of YiMMOgène. The OD 600 was measured in a micro-plate reader. ♦: OD 600 after 6 hour culture (early exponential phase). ●: OD 600 after 17 hour culture (late exponential phase). ▄: OD 600 after 24 hour culture (early stationary phase). C ) Percentage amplification of MNN5 from 47 YiMMOgène after different culture times in micro-plate. All amplifications have been carried out with the Platinium Taq.
    Figure Legend Snippet: Efficiency of MNN5 gene amplification from liquid culture of modified strain. A ) Percentage of MNN5 amplification from 47 colonies picked on Petri dishes and 47 colonies picked from liquid cultures in micro-plates. B ) Growth curve in micro-plate (YPD medium), average of two colonies of YiMMOgène. The OD 600 was measured in a micro-plate reader. ♦: OD 600 after 6 hour culture (early exponential phase). ●: OD 600 after 17 hour culture (late exponential phase). ▄: OD 600 after 24 hour culture (early stationary phase). C ) Percentage amplification of MNN5 from 47 YiMMOgène after different culture times in micro-plate. All amplifications have been carried out with the Platinium Taq.

    Techniques Used: Amplification, Modification

    MNN5 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 47 colonies for each condition. Dark grey: Petri dishes cultures, light grey: micro-plate cultures. B ) Average of the 3 assays presented in A (standard deviation =22.07). C) PCR product visualised on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications were carried out using Platinium Taq.
    Figure Legend Snippet: MNN5 gene amplification efficiency on modified strain colonies picked from Petri dishes or micro-plate cultures. A ) 3 independent assays carried out on 47 colonies for each condition. Dark grey: Petri dishes cultures, light grey: micro-plate cultures. B ) Average of the 3 assays presented in A (standard deviation =22.07). C) PCR product visualised on a 1% agarose gel stained with SYBR safe. Left panel (1): Petri dishes, right panel (2): micro-plate cultures, ML: Molecular ladder. All amplifications were carried out using Platinium Taq.

    Techniques Used: Amplification, Modification, Standard Deviation, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining

    Related Articles

    Clone Assay:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The PCR products containing bisulfite-resistant cytosines were ligated into the pCR2.1 vector (Invitrogen), and several clones were sequenced to confirm the presence of the amplified DNA.

    Amplification:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The reaction conditions consisted of 30 cycles of 95 °C for 1 min, 55 °C for 1 min and 74 °C for 2 min. A semi-nested PCR using the amplified products at a 1:50 dilution, the internal primer ML561 and primer MU379 was performed using similar PCR conditions as described above.

    Article Title: Novel Algorithms Reveal Streptococcal Transcriptomes and Clues about Undefined Genes
    Article Snippet: RT–PCR generation of amplicons was performed with the SuperScript III One-Step RT–PCR system with Platinum Taq DNA polymerase (Invitrogen) in reaction mixtures (50 μl) containing 0.2 μM of each gene-specific forward and reverse primers and 0.1 μg of DNase-treated, purified total RNA from late-log phase cultures (OD = 0.7) of strain SF370. .. RNA was converted to cDNA (50 °C for 30 min), which was then PCR amplified in the same tube (45 cycles of the following conditions: 94 °C for 15 s, 52 °C for 30 s, and 68 °C for 2 min).

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Even though extraction and amplification controls (1 per 7 samples) were consistently negative, we suspected that contamination of a PCR reagent might cause the lack of reproducibility and the consistent positivity rate of 5%. .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    Article Title: Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments
    Article Snippet: .. To enrich for circularized DNA by degrading linear strands including selectors, 10 μl of the circularization mixtures (0.4 μg DNA) were then added to a 10 μl mixture of 5 U Exonuclease I (New England Biolabs), 110 mM Tris–HCl, pH 9.0, 3 mM MgCl2 and 0.2 μg BSA and incubated for 2 h at 37°C, followed by 95°C for 10 min. Amplification was performed using 4 μl of each exonuclease-treated circularization reaction (80 ng DNA each) added to 17 μl mixture of 1× PCR buffer (Invitrogen), supplemented with 0.5 U Platinum Taq DNA polymerase (Invitrogen), 0.25 mM dNTP, 0.4 μM Cy-3-labeled forward and reverse primer, respectively, and 2 mM MgCl2 . .. Array hybridization cDNA arrays were obtained from the microarray core facility at Uppsala University.

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: .. Many researchers have since obtained successful amplification and improved results by substituting Invitrogen Platinum Taq DNA Polymerase for the standard Invitrogen Taq DNA Polymerase (e.g., [ ]). ..

    Article Title: Comparative Methods to Improve the Detection of BRAF V600 Mutations in Highly Pigmented Melanoma Specimens
    Article Snippet: .. Conventional PCR amplification was performed in a volume of 50 μl containing 1× PCR buffer, 10 μM deoxyribonucleoside triphosphate (dNTP), 10 μM each of the forward and reverse primers, 1 unit of Platinum Taq DNA polymerase (Life Technologies, Darmstadt, Germany), and 20 ng of genomic DNA. .. High-resolution melting (HRM) analysis PCR amplification and HRM analysis were performed on a Rotor-Gene 6000 (Corbett Research, Mortlake, Australia) by using the LightCycler 480 HRM Master Mix Kit (Roche Diagnostics, Meylan, France).

    Polymerase Chain Reaction:

    Article Title: Melt Analysis of Mismatch Amplification Mutation Assays (Melt-MAMA): A Functional Study of a Cost-Effective SNP Genotyping Assay in Bacterial Models
    Article Snippet: .. The conventional PCR master mix comprised two forward AS primers and a common reverse primer starting at 0.2 µM (IDT, San Diego, CA), 1x PCR buffer without MgCl2 (Invitrogen, Carlsbad, CA), 2 mM MgCl2 (Invitrogen, Carlsbad, CA), 200 µM of each dNTPs (Invitrogen, Carlsbad, CA), 0.8 units of Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 1 µl of template at ∼1ng/µl, and molecular grade water to a final volume of 10 µl. .. Agarose-MAMAs were performed under the following conditions: PCR amplifications were conducted on a MJ Research 96 well block thermal cycler DNA engines equipped with hot bonnets.

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: .. Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories). .. Melt curve analysis was included to assure a single PCR product was formed.

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The reaction conditions consisted of 30 cycles of 95 °C for 1 min, 55 °C for 1 min and 74 °C for 2 min. A semi-nested PCR using the amplified products at a 1:50 dilution, the internal primer ML561 and primer MU379 was performed using similar PCR conditions as described above.

    Article Title: Novel Algorithms Reveal Streptococcal Transcriptomes and Clues about Undefined Genes
    Article Snippet: RT–PCR generation of amplicons was performed with the SuperScript III One-Step RT–PCR system with Platinum Taq DNA polymerase (Invitrogen) in reaction mixtures (50 μl) containing 0.2 μM of each gene-specific forward and reverse primers and 0.1 μg of DNase-treated, purified total RNA from late-log phase cultures (OD = 0.7) of strain SF370. .. RNA was converted to cDNA (50 °C for 30 min), which was then PCR amplified in the same tube (45 cycles of the following conditions: 94 °C for 15 s, 52 °C for 30 s, and 68 °C for 2 min).

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Even though extraction and amplification controls (1 per 7 samples) were consistently negative, we suspected that contamination of a PCR reagent might cause the lack of reproducibility and the consistent positivity rate of 5%. .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    Article Title: Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments
    Article Snippet: .. To enrich for circularized DNA by degrading linear strands including selectors, 10 μl of the circularization mixtures (0.4 μg DNA) were then added to a 10 μl mixture of 5 U Exonuclease I (New England Biolabs), 110 mM Tris–HCl, pH 9.0, 3 mM MgCl2 and 0.2 μg BSA and incubated for 2 h at 37°C, followed by 95°C for 10 min. Amplification was performed using 4 μl of each exonuclease-treated circularization reaction (80 ng DNA each) added to 17 μl mixture of 1× PCR buffer (Invitrogen), supplemented with 0.5 U Platinum Taq DNA polymerase (Invitrogen), 0.25 mM dNTP, 0.4 μM Cy-3-labeled forward and reverse primer, respectively, and 2 mM MgCl2 . .. Array hybridization cDNA arrays were obtained from the microarray core facility at Uppsala University.

    Article Title: Epigenetic regulation of CpG promoter methylation in invasive prostate cancer cells
    Article Snippet: .. PCR was performed using Platinum Taq Polymerase (Invitrogen) and 200 ng of either genomic or bisulfite treated DNA. ..

    Article Title: Selective control of primer usage in multiplex one-step reverse transcription PCR
    Article Snippet: .. Reaction conditions included 1× PCR buffer (20 mM Tris (pH 8.4), 50 mM KCl) (Invitrogen), 1.5 mM MgCl2 (Invitrogen), gene-specific PCR primers (0.5 μM) (TriLink), oligo(dT)18 primer (1 μM) (TriLink), 0.16 mM dNTPs (New England Biolabs), 0.5 μL of human trachea total RNA or other human total RNA (Stratagene or Ambion, each at ~1.6 μg/μL), 5 U RNase Inhibitor (New England Biolabs), 50 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Invitrogen) or SuperScript® III Reverse Transcriptase (SSIII RT) (Invitrogen), and 0.6 U of Taq DNA Polymerase, recombinant (Invitrogen) or Platinum® Taq DNA Polymerase (Invitrogen) or AmpliTaq Gold® DNA Polymerase (Applied Biosystems), in a 50 μL reaction volume. .. Thermal cycling conditions utilized a reverse transcription step at 42°C for 30 min when M-MLV RT was used and 55°C for 30 min when SSIII RT was employed; 95°C for 10 min (RT inactivation and initial denaturation step), followed by 45 PCR cycles at 95°C for 30 sec, 60°C for 1 min and final extension at 72°C for 5 min.

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: Difficulties and biases in PCR amplification have been previously reported for the AR gene (e.g., [ ]), most likely due to the high GC content in many exonic trinucleotide repeat fragments including AR . .. Many researchers have since obtained successful amplification and improved results by substituting Invitrogen Platinum Taq DNA Polymerase for the standard Invitrogen Taq DNA Polymerase (e.g., [ ]).

    Article Title: Comparative Methods to Improve the Detection of BRAF V600 Mutations in Highly Pigmented Melanoma Specimens
    Article Snippet: .. Conventional PCR amplification was performed in a volume of 50 μl containing 1× PCR buffer, 10 μM deoxyribonucleoside triphosphate (dNTP), 10 μM each of the forward and reverse primers, 1 unit of Platinum Taq DNA polymerase (Life Technologies, Darmstadt, Germany), and 20 ng of genomic DNA. .. High-resolution melting (HRM) analysis PCR amplification and HRM analysis were performed on a Rotor-Gene 6000 (Corbett Research, Mortlake, Australia) by using the LightCycler 480 HRM Master Mix Kit (Roche Diagnostics, Meylan, France).

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: The presence of mouse DNA in PCR reagents emphasizes the critical importance of proper controls and carefully chosen, sensitive assays to detect trace amounts of mouse DNA. .. Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV.

    Blocking Assay:

    Article Title: Melt Analysis of Mismatch Amplification Mutation Assays (Melt-MAMA): A Functional Study of a Cost-Effective SNP Genotyping Assay in Bacterial Models
    Article Snippet: The conventional PCR master mix comprised two forward AS primers and a common reverse primer starting at 0.2 µM (IDT, San Diego, CA), 1x PCR buffer without MgCl2 (Invitrogen, Carlsbad, CA), 2 mM MgCl2 (Invitrogen, Carlsbad, CA), 200 µM of each dNTPs (Invitrogen, Carlsbad, CA), 0.8 units of Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 1 µl of template at ∼1ng/µl, and molecular grade water to a final volume of 10 µl. .. Agarose-MAMAs were performed under the following conditions: PCR amplifications were conducted on a MJ Research 96 well block thermal cycler DNA engines equipped with hot bonnets.

    SYBR Green Assay:

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: .. Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories). .. Melt curve analysis was included to assure a single PCR product was formed.

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA). .. SYBR Green® I nucleic acid stain was purchased from Invitrogen and passive reference ROX dye (1 mM) was purchased from Stratagene (La Jolla, CA, USA).

    Incubation:

    Article Title: Multiplex amplification enabled by selective circularization of large sets of genomic DNA fragments
    Article Snippet: .. To enrich for circularized DNA by degrading linear strands including selectors, 10 μl of the circularization mixtures (0.4 μg DNA) were then added to a 10 μl mixture of 5 U Exonuclease I (New England Biolabs), 110 mM Tris–HCl, pH 9.0, 3 mM MgCl2 and 0.2 μg BSA and incubated for 2 h at 37°C, followed by 95°C for 10 min. Amplification was performed using 4 μl of each exonuclease-treated circularization reaction (80 ng DNA each) added to 17 μl mixture of 1× PCR buffer (Invitrogen), supplemented with 0.5 U Platinum Taq DNA polymerase (Invitrogen), 0.25 mM dNTP, 0.4 μM Cy-3-labeled forward and reverse primer, respectively, and 2 mM MgCl2 . .. Array hybridization cDNA arrays were obtained from the microarray core facility at Uppsala University.

    Activity Assay:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV. .. This is not too surprising because the mouse monoclonal antibody used to prevent enzyme activity prior to heat activation might be the source of mouse DNA in the enzyme.

    Mass Spectrometry:

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: Electrospray mass spectrometry analyses were done by HT Laboratories (San Diego, CA, USA). .. Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Modification:

    Article Title: Epigenetic regulation of CpG promoter methylation in invasive prostate cancer cells
    Article Snippet: Methylation Specific polymerase chain reaction (MSP-PCR) A total of 1 μg of DNA extracted from total (parental) DU145 and LNCaP cells was bisulfite modified using the EpiTect Bisulfite kit from Qiagen. .. PCR was performed using Platinum Taq Polymerase (Invitrogen) and 200 ng of either genomic or bisulfite treated DNA.

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: Primers which contained either one PTE modification to the 3′-terminal internucleotide linkage or two modifications to the 3′-terminal and penultimate internucleotide linkages were also prepared by TriLink, on an ABI Expedite 8909 DNA synthesizer using standard manufacturer-suggested procedures as detailed in the Supplementary Data section. .. Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: .. Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories). .. Melt curve analysis was included to assure a single PCR product was formed.

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: We next tested each component of the nested PCR using the IAP qPCR assay in replicates of 8. .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    High Performance Liquid Chromatography:

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: HPLC was accomplished on a Beckman, Inc. (Fullerton, CA, USA) System Gold Nouveau Model 126 with Model 168 photodiode array detector. .. Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Concentration Assay:

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: From quantification, samples were normalized to a working concentration of 2.5 ng/ul and amplified with the primers developed by [ ], which capture a ~700 bp region of exon 1 containing three trinucleotide repeat tracts. .. Many researchers have since obtained successful amplification and improved results by substituting Invitrogen Platinum Taq DNA Polymerase for the standard Invitrogen Taq DNA Polymerase (e.g., [ ]).

    Viral Replication Assay:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: This made the viral replication assay very time-consuming and labor-intensive, and we could perform it only on a subset of our samples. .. Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV.

    Sequencing:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: To obtain products for sequencing, the converted DNA was amplified using semi-nested PCR as previously described by Dammann et al. [ ]. .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA).

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Sequencing these products revealed MLV-related sequences with 95 to 100% similarity to sequences published previously ( ; data not shown). .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    Recombinant:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences. ..

    Article Title: Selective control of primer usage in multiplex one-step reverse transcription PCR
    Article Snippet: .. Reaction conditions included 1× PCR buffer (20 mM Tris (pH 8.4), 50 mM KCl) (Invitrogen), 1.5 mM MgCl2 (Invitrogen), gene-specific PCR primers (0.5 μM) (TriLink), oligo(dT)18 primer (1 μM) (TriLink), 0.16 mM dNTPs (New England Biolabs), 0.5 μL of human trachea total RNA or other human total RNA (Stratagene or Ambion, each at ~1.6 μg/μL), 5 U RNase Inhibitor (New England Biolabs), 50 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Invitrogen) or SuperScript® III Reverse Transcriptase (SSIII RT) (Invitrogen), and 0.6 U of Taq DNA Polymerase, recombinant (Invitrogen) or Platinum® Taq DNA Polymerase (Invitrogen) or AmpliTaq Gold® DNA Polymerase (Applied Biosystems), in a 50 μL reaction volume. .. Thermal cycling conditions utilized a reverse transcription step at 42°C for 30 min when M-MLV RT was used and 55°C for 30 min when SSIII RT was employed; 95°C for 10 min (RT inactivation and initial denaturation step), followed by 45 PCR cycles at 95°C for 30 sec, 60°C for 1 min and final extension at 72°C for 5 min.

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV. .. What was surprising, however, was our finding mouse sequences in Invitrogen's recombinant Taq polymerase that is expressed in Escherichia coli ; we are not sure what the source of mouse DNA is, in this case.

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: .. Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA). .. DyNAzyme™ II Hot Start DNA polymerase and the deoxynucleotide solution set (dNTPs) were purchased from New England Biolabs (Ipswich, MA, USA).

    DNA Extraction:

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: Paragraph title: DNA extraction, quantification and amplification ... Many researchers have since obtained successful amplification and improved results by substituting Invitrogen Platinum Taq DNA Polymerase for the standard Invitrogen Taq DNA Polymerase (e.g., [ ]).

    Methylation:

    Article Title: Epigenetic regulation of CpG promoter methylation in invasive prostate cancer cells
    Article Snippet: Paragraph title: Methylation Specific polymerase chain reaction (MSP-PCR) ... PCR was performed using Platinum Taq Polymerase (Invitrogen) and 200 ng of either genomic or bisulfite treated DNA.

    Isolation:

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: Total RNA was isolated using RNeasy minikit (Qiagen, Venlo, The Netherlands) according to manufacturer's instructions. .. Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories).

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: Genomic DNA was isolated from the treated cells using the QIAamp DNA Mini Kit (Qiagen, Valencia, CA, USA). .. Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA).

    Size-exclusion Chromatography:

    Article Title: Melt Analysis of Mismatch Amplification Mutation Assays (Melt-MAMA): A Functional Study of a Cost-Effective SNP Genotyping Assay in Bacterial Models
    Article Snippet: The conventional PCR master mix comprised two forward AS primers and a common reverse primer starting at 0.2 µM (IDT, San Diego, CA), 1x PCR buffer without MgCl2 (Invitrogen, Carlsbad, CA), 2 mM MgCl2 (Invitrogen, Carlsbad, CA), 200 µM of each dNTPs (Invitrogen, Carlsbad, CA), 0.8 units of Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 1 µl of template at ∼1ng/µl, and molecular grade water to a final volume of 10 µl. .. PCRs were raised to 94°C for 5 min to denature the DNA and to activate the hot-start Taq DNA polymerase, then cycled at 94°C for 30 sec, 55°C–62.5°C (depended on assay) for 30 sec, 72°C for 30 sec, with a final extension at 72°C for 5 min.

    Article Title: Selective control of primer usage in multiplex one-step reverse transcription PCR
    Article Snippet: Reaction conditions included 1× PCR buffer (20 mM Tris (pH 8.4), 50 mM KCl) (Invitrogen), 1.5 mM MgCl2 (Invitrogen), gene-specific PCR primers (0.5 μM) (TriLink), oligo(dT)18 primer (1 μM) (TriLink), 0.16 mM dNTPs (New England Biolabs), 0.5 μL of human trachea total RNA or other human total RNA (Stratagene or Ambion, each at ~1.6 μg/μL), 5 U RNase Inhibitor (New England Biolabs), 50 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Invitrogen) or SuperScript® III Reverse Transcriptase (SSIII RT) (Invitrogen), and 0.6 U of Taq DNA Polymerase, recombinant (Invitrogen) or Platinum® Taq DNA Polymerase (Invitrogen) or AmpliTaq Gold® DNA Polymerase (Applied Biosystems), in a 50 μL reaction volume. .. Thermal cycling conditions utilized a reverse transcription step at 42°C for 30 min when M-MLV RT was used and 55°C for 30 min when SSIII RT was employed; 95°C for 10 min (RT inactivation and initial denaturation step), followed by 45 PCR cycles at 95°C for 30 sec, 60°C for 1 min and final extension at 72°C for 5 min.

    Labeling:

    Article Title: A test of somatic mosaicism in the androgen receptor gene of Canada lynx (Lynx canadensis)
    Article Snippet: Amplification was conducted in a 10ul reaction containing deionized water (Invitrogen), 1X PCR Reaction Buffer (Invitrogen), 2 mM MgCl2 (Invitrogen), 0.2 mM dNTP solution (Invitrogen), 0.2 mg/mL BSA, 0.4uM forward and reverse primers (forward primer labeled with the fluorescent dye HEX) (Integrated DNA Technologies), 0.025U Invitrogen Platinum Taq DNA Polymerase, and 5 ng of DNA. .. Many researchers have since obtained successful amplification and improved results by substituting Invitrogen Platinum Taq DNA Polymerase for the standard Invitrogen Taq DNA Polymerase (e.g., [ ]).

    Purification:

    Article Title: Novel Algorithms Reveal Streptococcal Transcriptomes and Clues about Undefined Genes
    Article Snippet: .. RT–PCR generation of amplicons was performed with the SuperScript III One-Step RT–PCR system with Platinum Taq DNA polymerase (Invitrogen) in reaction mixtures (50 μl) containing 0.2 μM of each gene-specific forward and reverse primers and 0.1 μg of DNase-treated, purified total RNA from late-log phase cultures (OD = 0.7) of strain SF370. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Novel Algorithms Reveal Streptococcal Transcriptomes and Clues about Undefined Genes
    Article Snippet: .. RT–PCR generation of amplicons was performed with the SuperScript III One-Step RT–PCR system with Platinum Taq DNA polymerase (Invitrogen) in reaction mixtures (50 μl) containing 0.2 μM of each gene-specific forward and reverse primers and 0.1 μg of DNase-treated, purified total RNA from late-log phase cultures (OD = 0.7) of strain SF370. ..

    Article Title: Selective control of primer usage in multiplex one-step reverse transcription PCR
    Article Snippet: Paragraph title: One-step RT-PCR (Endpoint) ... Reaction conditions included 1× PCR buffer (20 mM Tris (pH 8.4), 50 mM KCl) (Invitrogen), 1.5 mM MgCl2 (Invitrogen), gene-specific PCR primers (0.5 μM) (TriLink), oligo(dT)18 primer (1 μM) (TriLink), 0.16 mM dNTPs (New England Biolabs), 0.5 μL of human trachea total RNA or other human total RNA (Stratagene or Ambion, each at ~1.6 μg/μL), 5 U RNase Inhibitor (New England Biolabs), 50 U Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) (Invitrogen) or SuperScript® III Reverse Transcriptase (SSIII RT) (Invitrogen), and 0.6 U of Taq DNA Polymerase, recombinant (Invitrogen) or Platinum® Taq DNA Polymerase (Invitrogen) or AmpliTaq Gold® DNA Polymerase (Applied Biosystems), in a 50 μL reaction volume.

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: .. Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV. .. This is not too surprising because the mouse monoclonal antibody used to prevent enzyme activity prior to heat activation might be the source of mouse DNA in the enzyme.

    Quantitative RT-PCR:

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: Paragraph title: Real-time RT-PCR ... Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories).

    Nested PCR:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Paragraph title: Mouse DNA is present in a reagent used in previously published nested-PCR assay. ... We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    Plasmid Preparation:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA). .. The PCR products containing bisulfite-resistant cytosines were ligated into the pCR2.1 vector (Invitrogen), and several clones were sequenced to confirm the presence of the amplified DNA.

    Irradiation:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: We prevented this by handling only one set of cultures in the biosafety cabinet at a time and meticulously decontaminated the cabinet between cultures with 70% ethanol and UV irradiation. .. Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV.

    Agarose Gel Electrophoresis:

    Article Title: Melt Analysis of Mismatch Amplification Mutation Assays (Melt-MAMA): A Functional Study of a Cost-Effective SNP Genotyping Assay in Bacterial Models
    Article Snippet: The conventional PCR master mix comprised two forward AS primers and a common reverse primer starting at 0.2 µM (IDT, San Diego, CA), 1x PCR buffer without MgCl2 (Invitrogen, Carlsbad, CA), 2 mM MgCl2 (Invitrogen, Carlsbad, CA), 200 µM of each dNTPs (Invitrogen, Carlsbad, CA), 0.8 units of Platinum Taq DNA polymerase (Invitrogen, Carlsbad, CA), 1 µl of template at ∼1ng/µl, and molecular grade water to a final volume of 10 µl. .. The PCR amplicons and a 100 bp ladder (Invitrogen, Carlsbad, CA) were electrophoresed at 100 V for 90–110 minutes on a 2% or 3% agarose gel (Fisher Scientific, Pittsburgh, PA) prepared in 1x TAE and visualized with SYBR Safe (Invitrogen, Carlsbad, CA).

    Article Title: Epigenetic regulation of CpG promoter methylation in invasive prostate cancer cells
    Article Snippet: PCR was performed using Platinum Taq Polymerase (Invitrogen) and 200 ng of either genomic or bisulfite treated DNA. .. A portion of the PCR product was run on a 1% agarose gel containing ethidum bromide.

    Nuclear Magnetic Resonance:

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: NMR spectra were recorded on Bruker Model AX 500 spectrometer (NuMega, San Diego, CA, USA). .. Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA).

    Spectrophotometry:

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids
    Article Snippet: RNA sample concentrations were determined using a NanoDrop ND-1000 spectrophotometer (Isogen, Maarssen, The Netherlands). .. Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories).

    DNA Methylation Assay:

    Article Title: Epigenetic Reactivation of RASSF1A by Phenethyl Isothiocyanate (PEITC) and promotion of apoptosis in LNCaP cells
    Article Snippet: Paragraph title: 2.2. DNA methylation analysis ... Briefly, the DNA sequences were amplified by mixing bisulfite-converted DNA (500 ng) with primer MU379 (50 pmoles) and primer ML730 (50 pmoles) in reaction buffer (20 μl) containing dNTPs (200 μM each dNTP) using Platinum PCR Taq DNA polymerase (Invitrogen, Carlsbad, CA, USA).

    Produced:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: We tested 36 replicates of genomic DNA from uninfected LNCaP cells with the nested PCR and found that 2 produced a positive result ( C, a subset of the data). .. We discovered that both recombinant Taq polymerase (Invitrogen) and the Platinum Taq polymerase (Invitrogen) tested positive for IAP sequences.

    Activation Assay:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV. .. This is not too surprising because the mouse monoclonal antibody used to prevent enzyme activity prior to heat activation might be the source of mouse DNA in the enzyme.

    Staining:

    Article Title: Novel Algorithms Reveal Streptococcal Transcriptomes and Clues about Undefined Genes
    Article Snippet: RT–PCR generation of amplicons was performed with the SuperScript III One-Step RT–PCR system with Platinum Taq DNA polymerase (Invitrogen) in reaction mixtures (50 μl) containing 0.2 μM of each gene-specific forward and reverse primers and 0.1 μg of DNase-treated, purified total RNA from late-log phase cultures (OD = 0.7) of strain SF370. .. Resulting DNA fragments were separated on 1% agarose gels in TAE buffer and visualized by ethidium bromide staining.

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: Thermus aquaticus (Taq ) DNA polymerase (recombinant) and Platinum® Taq DNA Polymerases were purchased from Invitrogen (Carlsbad, CA, USA). .. SYBR Green® I nucleic acid stain was purchased from Invitrogen and passive reference ROX dye (1 mM) was purchased from Stratagene (La Jolla, CA, USA).

    Hood:

    Article Title: Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿Absence of XMRV Retrovirus and Other Murine Leukemia Virus-Related Viruses in Patients with Chronic Fatigue Syndrome ▿ ¶
    Article Snippet: We prevented this by handling only one set of cultures in the biosafety cabinet at a time and meticulously decontaminated the cabinet between cultures with 70% ethanol and UV irradiation. .. Sato et al. , using a sensitive RT-PCR kit, found that Platinum Taq polymerase (Invitrogen) contained RNA from polytropic endogenous MLV.

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    Thermo Fisher platinum taq dna polymerase high fidelity
    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by <t>DNA</t> sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the <t>Taq</t> DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.
    Platinum Taq Dna Polymerase High Fidelity, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 417 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/platinum taq dna polymerase high fidelity/product/Thermo Fisher
    Average 99 stars, based on 417 article reviews
    Price from $9.99 to $1999.99
    platinum taq dna polymerase high fidelity - by Bioz Stars, 2020-04
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    99
    Thermo Fisher superscript iii rt pcr platinum taq high fidelity kit
    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all <t>three</t> of the cell populations were subjected to <t>RT-PCR</t> to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).
    Superscript Iii Rt Pcr Platinum Taq High Fidelity Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii rt pcr platinum taq high fidelity kit/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Journal: BioMed Research International

    Article Title: LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity

    doi: 10.1155/2018/5657085

    Figure Lengend Snippet: Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Article Snippet: PCR fragments of LraI operon amplified using Platinum™ Taq DNA Polymerase High Fidelity were cloned into pAZIL vector predigested with Sma I.

    Techniques: DNA Sequencing, Sequencing, Activity Assay

    Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Journal: BioMed Research International

    Article Title: LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity

    doi: 10.1155/2018/5657085

    Figure Lengend Snippet: Determination of the Lra I cleavage site 5′-G/AATTC-3′ of pBluescipt SK+ by DNA sequencing. (a) Sequence of pBluescipt SK+ predigested with Lra I restriction enzyme and (b) with Eco RI obtained using M13R primer. The colour-coded sequence traces are A (green), T (red), C (blue), and G (black). The extra A base was added at the end of the cleaved template by the Taq DNA polymerase used for sequencing due to template-independent terminal nucleotide transferase activity of Taq DNA polymerase.

    Article Snippet: Platinum™ Taq DNA Polymerase High Fidelity (Thermo Fisher Scientific, Waltham, MA, USA) was used to amplify DNA fragments by PCR in GeneAmp PCR system 2700 thermal cycler (Applied Biosystems, Foster City, CA, USA).

    Techniques: DNA Sequencing, Sequencing, Activity Assay

    ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Journal: Journal of Virology

    Article Title: Transposon Mutagenesis of the Zika Virus Genome Highlights Regions Essential for RNA Replication and Restricted for Immune Evasion

    doi: 10.1128/JVI.00698-17

    Figure Lengend Snippet: ZIKV insertional mutant library screening protocol and deep sequencing results. (A) Representative images of individual mutants from the library of ZIKV cDNA clones (blue), each bearing a single 15-nucleotide insertion, represented by a colored line (not drawn to scale). (B) 293T cells were transfected with the mutant plasmid library to select for mutant genomes that could initiate RNA replication and virus release. (C) Forty-eight hours after transfection, the supernatant was collected and passaged on Vero cells to select mutants that retained the ability to spread. (D) At 48 h postinfection, the supernatant was passaged once more on Vero cells to select for the fittest mutants. At the time of supernatant collection, RNAs from all three of the cell populations were subjected to RT-PCR to amplify the viral genomes. The subsequent RT-PCR products were then deep sequenced. (E) Raw number of deep sequence reads with inserts at each location for the input plasmid library. The ZIKV genome schematic and bars in the graph show structural genes in red, nonstructural genes in blue, UTRs in black, and the intron used to stabilize the plasmid in bacteria in yellow. This intron was not present in the rescued viruses. Alternating gray and white shading within each graph differentiates the genomic regions. (F to H) Frequencies of insertions at each location following normalization to the representation in the input library after plasmid DNA transfection (F), infected cell passage 1 (G), and infected cell passage 2 (H), with a schematic of the genome shown below. Numbering along the x axis indicates the nucleotide position in the ZIKV genome, while the y axis gives the log raw number of reads (E) or the log fold change over input (F to H), with horizontal dotted lines representing the cutoff of the mutants enriched two standard deviations above the average. Values are averages for three independent screens (see Materials and Methods for details).

    Article Snippet: The viral RNA was then amplified in three segments by utilizing a SuperScript III RT-PCR Platinum Taq High Fidelity kit (Thermo Fisher Scientific).

    Techniques: Mutagenesis, Library Screening, Sequencing, Transfection, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Infection