t4 dna polymerase  (Roche)


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    Structured Review

    Roche t4 dna polymerase
    T4 Dna Polymerase, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna polymerase/product/Roche
    Average 92 stars, based on 33 article reviews
    Price from $9.99 to $1999.99
    t4 dna polymerase - by Bioz Stars, 2020-09
    92/100 stars

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    Mutagenesis:

    Article Title: Mutations in the Stalk of the Measles Virus Hemagglutinin Protein Decrease Fusion but Do Not Interfere with Virus-Specific Interaction with the Homologous Fusion Protein ▿
    Article Snippet: .. Mutagenesis primers (Integrated DNA Technologies, Coralville, IA) were annealed to the template and extended with T4 DNA polymerase, and the ends were ligated with T4 DNA ligase (Roche). .. The mutagenesis reaction products were transformed into MV1190 cells (Bio-Rad, Hercules, CA) that were then selected for ampicillin resistance.

    IA:

    Article Title: Mutations in the Stalk of the Measles Virus Hemagglutinin Protein Decrease Fusion but Do Not Interfere with Virus-Specific Interaction with the Homologous Fusion Protein ▿
    Article Snippet: .. Mutagenesis primers (Integrated DNA Technologies, Coralville, IA) were annealed to the template and extended with T4 DNA polymerase, and the ends were ligated with T4 DNA ligase (Roche). .. The mutagenesis reaction products were transformed into MV1190 cells (Bio-Rad, Hercules, CA) that were then selected for ampicillin resistance.

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  • 94
    Roche dnase i
    DNase I-PCR Assay of the <t>DNase</t> I Sensitivity of a Transgenic Enhancer Sequence. (A) . (B) A diagram illustrating the design of a PCR primer that allows the transgenic L3 locus, but not the endogenous L3, to be specifically amplified. (C) DNase I-PCR shows differential DNase I sensitivity of chromatin in selected genomic regions. The DNase I sensitivity of the transgenic L3 locus was similar to that of the open chromatin control ACTIN7 .
    Dnase I, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 650 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dnase i/product/Roche
    Average 94 stars, based on 650 article reviews
    Price from $9.99 to $1999.99
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    94/100 stars
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    94
    Roche rnase free dnase i
    Analysis of transcript levels in 7 h germlings. A. parasiticus SU-1, AFS10, and Δ veA were grown on YES solid media and spores were collected at 5 days. Germlings were grown from fresh spores in GMS liquid medium for 7 h, frozen in liquid nitrogen, and stored at −80 °C. RNA was extracted from germlings using grinding and sonication as described in Methods. RT-PCR was performed on total RNA treated with <t>RNAse-free</t> <t>DNAse</t> I with primers specific for each gene ( Table 3 ). PCR products were separated on a 1% agarose gel by electrophoresis. Citrate synthase, a constitutively expressed gene, was used as a positive control. gDNA, genomic DNA.
    Rnase Free Dnase I, supplied by Roche, used in various techniques. Bioz Stars score: 94/100, based on 358 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rnase free dnase i/product/Roche
    Average 94 stars, based on 358 article reviews
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    86
    Roche t4 dna polymerase treated insert
    Cloning strategy. The vector contains two appropriately oriented BsaI sites (A) upon digestion with BsaI linearized vector is obtained with ends having 4-base 5′-overhangs (B) shown in red. The recognition sequence of restriction enzyme BsaI are underlined and the cleavage site is marked. The Gene Of Interest (GOI) is amplified using two gene-specific primers with 7-base long additional sequence at the 5′ end (C) shown in bold. Treatment of PCR product with <t>T4</t> DNA polymerase and dTTP produces two different four-base overhangs that are complementary to two ends of the linearized vector shown in red (D). The ligation results in direction cloning of the insert into the vector (E).
    T4 Dna Polymerase Treated Insert, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 86 stars, based on 1 article reviews
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    DNase I-PCR Assay of the DNase I Sensitivity of a Transgenic Enhancer Sequence. (A) . (B) A diagram illustrating the design of a PCR primer that allows the transgenic L3 locus, but not the endogenous L3, to be specifically amplified. (C) DNase I-PCR shows differential DNase I sensitivity of chromatin in selected genomic regions. The DNase I sensitivity of the transgenic L3 locus was similar to that of the open chromatin control ACTIN7 .

    Journal: The Plant Cell

    Article Title: Genome-Wide Prediction and Validation of Intergenic Enhancers in Arabidopsis Using Open Chromatin Signatures [OPEN]

    doi: 10.1105/tpc.15.00537

    Figure Lengend Snippet: DNase I-PCR Assay of the DNase I Sensitivity of a Transgenic Enhancer Sequence. (A) . (B) A diagram illustrating the design of a PCR primer that allows the transgenic L3 locus, but not the endogenous L3, to be specifically amplified. (C) DNase I-PCR shows differential DNase I sensitivity of chromatin in selected genomic regions. The DNase I sensitivity of the transgenic L3 locus was similar to that of the open chromatin control ACTIN7 .

    Article Snippet: A DNase I (RNase-free; 10 U/μL; Roche Applied Science; catalog number 04716728001) dilution series was prepared by step-wise dilution using digestion buffer.

    Techniques: Polymerase Chain Reaction, Transgenic Assay, Sequencing, Amplification

    Influence of DNase I dosages on the sensitivity of PCR assays. Rate of positive PCR results at the detection limit are given depending on DNase I dosages for assays I (A), II (B), and III (C) (for details, see text). The threshold dosage of DNase I that was required for the complete elimination of false-positive PCR results is underlined. Amounts of DNase I increasing stepwise reduced the sensitivity of the PCRs.

    Journal: Journal of Clinical Microbiology

    Article Title: DNase Pretreatment of Master Mix Reagents Improves the Validity of Universal 16S rRNA Gene PCR Results

    doi: 10.1128/JCM.41.4.1763-1765.2003

    Figure Lengend Snippet: Influence of DNase I dosages on the sensitivity of PCR assays. Rate of positive PCR results at the detection limit are given depending on DNase I dosages for assays I (A), II (B), and III (C) (for details, see text). The threshold dosage of DNase I that was required for the complete elimination of false-positive PCR results is underlined. Amounts of DNase I increasing stepwise reduced the sensitivity of the PCRs.

    Article Snippet: However, when the master mix was pretreated with 25 IU of DNase I, the same amount of template gave a positive result in only 2 of 14 (14%) valid runs.

    Techniques: Polymerase Chain Reaction

    DNase I dosages required to improve universal PCR results. Percentages of PCR runs delivering valid (white bars), false-positive (grey bars), and false-negative (black bars) results depending on DNase I dosages are shown for assays I (A), II (B), and III (C) (for details, see text). Complete elimination of false-positive results and significant increases in valid results were achieved with highly varying amounts of DNase I (0.1 through 70 IU). The asterisks mark the significantly high proportion of valid PCR results obtained with DNase pretreatment compared to the results from PCR runs without DNase pretreatment.

    Journal: Journal of Clinical Microbiology

    Article Title: DNase Pretreatment of Master Mix Reagents Improves the Validity of Universal 16S rRNA Gene PCR Results

    doi: 10.1128/JCM.41.4.1763-1765.2003

    Figure Lengend Snippet: DNase I dosages required to improve universal PCR results. Percentages of PCR runs delivering valid (white bars), false-positive (grey bars), and false-negative (black bars) results depending on DNase I dosages are shown for assays I (A), II (B), and III (C) (for details, see text). Complete elimination of false-positive results and significant increases in valid results were achieved with highly varying amounts of DNase I (0.1 through 70 IU). The asterisks mark the significantly high proportion of valid PCR results obtained with DNase pretreatment compared to the results from PCR runs without DNase pretreatment.

    Article Snippet: However, when the master mix was pretreated with 25 IU of DNase I, the same amount of template gave a positive result in only 2 of 14 (14%) valid runs.

    Techniques: Polymerase Chain Reaction

    Analysis of transcript levels in 7 h germlings. A. parasiticus SU-1, AFS10, and Δ veA were grown on YES solid media and spores were collected at 5 days. Germlings were grown from fresh spores in GMS liquid medium for 7 h, frozen in liquid nitrogen, and stored at −80 °C. RNA was extracted from germlings using grinding and sonication as described in Methods. RT-PCR was performed on total RNA treated with RNAse-free DNAse I with primers specific for each gene ( Table 3 ). PCR products were separated on a 1% agarose gel by electrophoresis. Citrate synthase, a constitutively expressed gene, was used as a positive control. gDNA, genomic DNA.

    Journal: Toxins

    Article Title: Aflatoxin Biosynthesis Is a Novel Source of Reactive Oxygen Species—A Potential Redox Signal to Initiate Resistance to Oxidative Stress?

    doi: 10.3390/toxins7051411

    Figure Lengend Snippet: Analysis of transcript levels in 7 h germlings. A. parasiticus SU-1, AFS10, and Δ veA were grown on YES solid media and spores were collected at 5 days. Germlings were grown from fresh spores in GMS liquid medium for 7 h, frozen in liquid nitrogen, and stored at −80 °C. RNA was extracted from germlings using grinding and sonication as described in Methods. RT-PCR was performed on total RNA treated with RNAse-free DNAse I with primers specific for each gene ( Table 3 ). PCR products were separated on a 1% agarose gel by electrophoresis. Citrate synthase, a constitutively expressed gene, was used as a positive control. gDNA, genomic DNA.

    Article Snippet: RT-PCR was performed on total RNA treated with RNAse-free DNAse I (Roche Diagnostics, Indianapolis, IN, USA) with primers specific for the coding region of each gene.

    Techniques: Sonication, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Electrophoresis, Positive Control

    Cloning strategy. The vector contains two appropriately oriented BsaI sites (A) upon digestion with BsaI linearized vector is obtained with ends having 4-base 5′-overhangs (B) shown in red. The recognition sequence of restriction enzyme BsaI are underlined and the cleavage site is marked. The Gene Of Interest (GOI) is amplified using two gene-specific primers with 7-base long additional sequence at the 5′ end (C) shown in bold. Treatment of PCR product with T4 DNA polymerase and dTTP produces two different four-base overhangs that are complementary to two ends of the linearized vector shown in red (D). The ligation results in direction cloning of the insert into the vector (E).

    Journal: PLoS ONE

    Article Title: Rapid Restriction Enzyme-Free Cloning of PCR Products: A High-Throughput Method Applicable for Library Construction

    doi: 10.1371/journal.pone.0111538

    Figure Lengend Snippet: Cloning strategy. The vector contains two appropriately oriented BsaI sites (A) upon digestion with BsaI linearized vector is obtained with ends having 4-base 5′-overhangs (B) shown in red. The recognition sequence of restriction enzyme BsaI are underlined and the cleavage site is marked. The Gene Of Interest (GOI) is amplified using two gene-specific primers with 7-base long additional sequence at the 5′ end (C) shown in bold. Treatment of PCR product with T4 DNA polymerase and dTTP produces two different four-base overhangs that are complementary to two ends of the linearized vector shown in red (D). The ligation results in direction cloning of the insert into the vector (E).

    Article Snippet: The contents were mixed well, centrifuged at 4°C and then placed in a thermocycler for incubation at 15°C for 60 min followed by heat inactivation at 75°C for 20 min. Ligation was performed in a total volume of 10 µl containing 1 µl of 10X ligation buffer (New England Biolabs), 25 ng BsaI-HF-digested vector, 1 µl of T4 DNA Polymerase-treated insert (2–5 fold molar excess relative to the vector) and 1.0 unit of T4 DNA ligase (Roche) for 16 h at 16°C followed by 1 h at 37°C and heat inactivation at 65°C for 10 min. For electroporation, the ligation mixture was diluted ten-fold in water and then 1 µl of the diluted ligation sample (containing 250 pg vector equivalent) was electroporated into 25 µl of E. coli competent cells [BL21 (DE3) RIL, Agilent Technologies, CA, USA; efficiency 3–5×108 /µg].

    Techniques: Clone Assay, Plasmid Preparation, Sequencing, Amplification, Polymerase Chain Reaction, Ligation