mcrbc restriction endonuclease  (New England Biolabs)


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    Name:
    McrBC
    Description:
    McrBC 2 500 units
    Catalog Number:
    m0272l
    Price:
    290
    Size:
    2 500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs mcrbc restriction endonuclease
    McrBC
    McrBC 2 500 units
    https://www.bioz.com/result/mcrbc restriction endonuclease/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mcrbc restriction endonuclease - by Bioz Stars, 2020-05
    99/100 stars

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    1) Product Images from "Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential"

    Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential

    Journal: BMC Genomics

    doi: 10.1186/s12864-016-2545-1

    qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent McrBC enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2
    Figure Legend Snippet: qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent McrBC enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2

    Techniques Used: Real-time Polymerase Chain Reaction, Microarray, DNA Methylation Assay, Expressing, Methylated DNA Immunoprecipitation, CpG Methylation Assay, Methylation, Binding Assay, Mouse Assay, Two Tailed Test

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential
    Article Snippet: .. Validation of CpG methylation CpG methylation levels were examined by using Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) with McrBC restriction endonuclease (NEB, cat. no. M0272). ..

    Article Title: Extraordinary transgressive phenotypes of hybrid tomato are influenced by epigenetics and small silencing RNAs
    Article Snippet: .. About 500 ng of DNA at 20 ng/μl was incubated with 20 U McrBC (New England Biolabs) or an equivalent volume of 50% glycerol for 2 h at 37°C followed by heat inactivation at 80°C for 20 min. Target regions were amplified by qPCR from 40-ng digested DNA using SYBR Green JumpStart Taq ReadyMix (Sigma S9194), using primers described in . .. Total DNA (500 ng) from parents and hybrids was treated with sodium bisulphite using EZ-DNA methylation gold kit (Zymo Research) as per manufacturer's instructions.

    Negative Control:

    Article Title: PbGA2ox8 induces vascular-related anthocyanin accumulation and contributes to red stripe formation on pear fruit
    Article Snippet: .. The isolated DNA was then digested with 40 units of the methylation-sensitive restriction enzyme McrBC (New England Biolabs; M0272L) for 2 h, and the digestion buffer without GTPase was used as the negative control. .. The methylation level in the digested DNA templates was measured by semiquantitative PCR.

    Article Title: High-resolution spatiotemporal transcriptome mapping of tomato fruit development and ripening
    Article Snippet: .. A total of 0.5 μg DNA was digested with 7.5 U McrBC (NEB) according to the manufacturer’s recommendations for 6 h at 37 °C, or the same reaction without GTP as a negative control. .. PCR was performed using 40 ng McrBC-treated DNA.

    Amplification:

    Article Title: Extraordinary transgressive phenotypes of hybrid tomato are influenced by epigenetics and small silencing RNAs
    Article Snippet: .. About 500 ng of DNA at 20 ng/μl was incubated with 20 U McrBC (New England Biolabs) or an equivalent volume of 50% glycerol for 2 h at 37°C followed by heat inactivation at 80°C for 20 min. Target regions were amplified by qPCR from 40-ng digested DNA using SYBR Green JumpStart Taq ReadyMix (Sigma S9194), using primers described in . .. Total DNA (500 ng) from parents and hybrids was treated with sodium bisulphite using EZ-DNA methylation gold kit (Zymo Research) as per manufacturer's instructions.

    Methylation:

    Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential
    Article Snippet: .. Validation of CpG methylation CpG methylation levels were examined by using Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) with McrBC restriction endonuclease (NEB, cat. no. M0272). ..

    Article Title: PbGA2ox8 induces vascular-related anthocyanin accumulation and contributes to red stripe formation on pear fruit
    Article Snippet: .. The isolated DNA was then digested with 40 units of the methylation-sensitive restriction enzyme McrBC (New England Biolabs; M0272L) for 2 h, and the digestion buffer without GTPase was used as the negative control. .. The methylation level in the digested DNA templates was measured by semiquantitative PCR.

    Article Title: Independent functions of DNMT1 and USP7 at replication foci
    Article Snippet: .. Genomic DNA was digested for two rounds with methylation-sensitive enzyme HpaII, its isoschizomer MspI as a control, or McrBC (all from NEB). .. DNA was quantified and ran on 0.8% agarose gel, which was stained with ethidium bromide.

    Article Title: Chromatin and sequence features that define the fine and gross structure of genomic methylation patterns
    Article Snippet: .. Unmethylated and methylated compartments were obtained by limit digestions of 10–15 μg of DNA with McrBC or five tetranucleotide methylation-sensitive restriction endonucleases (referred to as RE), respectively (New England BioLabs). ..

    Isolation:

    Article Title: PbGA2ox8 induces vascular-related anthocyanin accumulation and contributes to red stripe formation on pear fruit
    Article Snippet: .. The isolated DNA was then digested with 40 units of the methylation-sensitive restriction enzyme McrBC (New England Biolabs; M0272L) for 2 h, and the digestion buffer without GTPase was used as the negative control. .. The methylation level in the digested DNA templates was measured by semiquantitative PCR.

    SYBR Green Assay:

    Article Title: Extraordinary transgressive phenotypes of hybrid tomato are influenced by epigenetics and small silencing RNAs
    Article Snippet: .. About 500 ng of DNA at 20 ng/μl was incubated with 20 U McrBC (New England Biolabs) or an equivalent volume of 50% glycerol for 2 h at 37°C followed by heat inactivation at 80°C for 20 min. Target regions were amplified by qPCR from 40-ng digested DNA using SYBR Green JumpStart Taq ReadyMix (Sigma S9194), using primers described in . .. Total DNA (500 ng) from parents and hybrids was treated with sodium bisulphite using EZ-DNA methylation gold kit (Zymo Research) as per manufacturer's instructions.

    Incubation:

    Article Title: Exendin-4 promotes extracellular-superoxide dismutase expression in A549 cells through DNA demethylation
    Article Snippet: .. Cleaved genomic DNA (500 ng) was further cleaved with McrBC (New England BioLabs, Beverly, MA), an endonuclease that cleaves DNA containing methylcytosine, in a final reaction volume of 10 µl at 37°C for 1 h followed by an incubation at 65°C for 20 min. .. The cleaved genomic DNA was diluted 10-fold with water, and 2 µl of the DNA was used as a template for the real-time RT-PCR analysis.

    Article Title: Extraordinary transgressive phenotypes of hybrid tomato are influenced by epigenetics and small silencing RNAs
    Article Snippet: .. About 500 ng of DNA at 20 ng/μl was incubated with 20 U McrBC (New England Biolabs) or an equivalent volume of 50% glycerol for 2 h at 37°C followed by heat inactivation at 80°C for 20 min. Target regions were amplified by qPCR from 40-ng digested DNA using SYBR Green JumpStart Taq ReadyMix (Sigma S9194), using primers described in . .. Total DNA (500 ng) from parents and hybrids was treated with sodium bisulphite using EZ-DNA methylation gold kit (Zymo Research) as per manufacturer's instructions.

    CpG Methylation Assay:

    Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential
    Article Snippet: .. Validation of CpG methylation CpG methylation levels were examined by using Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) with McrBC restriction endonuclease (NEB, cat. no. M0272). ..

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    New England Biolabs mcrbc restriction endonuclease
    qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative <t>PCR</t> (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent <t>McrBC</t> enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2
    Mcrbc Restriction Endonuclease, supplied by New England Biolabs, 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/mcrbc restriction endonuclease/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mcrbc restriction endonuclease - by Bioz Stars, 2020-05
    99/100 stars
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    qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent McrBC enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2

    Journal: BMC Genomics

    Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential

    doi: 10.1186/s12864-016-2545-1

    Figure Lengend Snippet: qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent McrBC enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2

    Article Snippet: Validation of CpG methylation CpG methylation levels were examined by using Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) with McrBC restriction endonuclease (NEB, cat. no. M0272).

    Techniques: Real-time Polymerase Chain Reaction, Microarray, DNA Methylation Assay, Expressing, Methylated DNA Immunoprecipitation, CpG Methylation Assay, Methylation, Binding Assay, Mouse Assay, Two Tailed Test