mnl i  (New England Biolabs)


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    Name:
    MnlI
    Description:
    MnlI 2 500 units
    Catalog Number:
    r0163l
    Price:
    282
    Size:
    2 500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs mnl i
    MnlI
    MnlI 2 500 units
    https://www.bioz.com/result/mnl i/product/New England Biolabs
    Average 98 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    mnl i - by Bioz Stars, 2020-09
    98/100 stars

    Images

    1) Product Images from "Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings"

    Article Title: Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2017.01683

    Assessment of RNF168 mutation (A) and RNF168 protein level (B) . (A) Direct sequencing of RNF168 exon 12 reveals homozygosity for the novel frameshift mutation c.295delG in genomic DNA from either of both patients (HA591, HA592). (B) RFLP analysis of PCR products on 2% agarose gel electrophoresis. S, size marker; 1, undigested PCR product; 2–6, PCR products cleaved with Mnl I: 2, wild-type control, 3, paternal sample, 4. maternal sample, 5, patient HA591, 6, patient HA592. (C) Western blot analysis reveals strongly reduced immunoreactivity for RNF168 protein in lymphoblastoid cells from either of both patients (HA591, HA592). Lymphoblastoid cell lines (LCLs) from a healthy individual were used as an RNF168-proficient control (HA325), and LCLs from a patient with classical ataxia–telangiectasia were used for comparison (HA56). β-actin served as the loading control (ACTB).
    Figure Legend Snippet: Assessment of RNF168 mutation (A) and RNF168 protein level (B) . (A) Direct sequencing of RNF168 exon 12 reveals homozygosity for the novel frameshift mutation c.295delG in genomic DNA from either of both patients (HA591, HA592). (B) RFLP analysis of PCR products on 2% agarose gel electrophoresis. S, size marker; 1, undigested PCR product; 2–6, PCR products cleaved with Mnl I: 2, wild-type control, 3, paternal sample, 4. maternal sample, 5, patient HA591, 6, patient HA592. (C) Western blot analysis reveals strongly reduced immunoreactivity for RNF168 protein in lymphoblastoid cells from either of both patients (HA591, HA592). Lymphoblastoid cell lines (LCLs) from a healthy individual were used as an RNF168-proficient control (HA325), and LCLs from a patient with classical ataxia–telangiectasia were used for comparison (HA56). β-actin served as the loading control (ACTB).

    Techniques Used: Mutagenesis, Sequencing, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker, Western Blot

    2) Product Images from "Coyotes (Canis latrans) as the Reservoir for a Human Pathogenic Bartonella sp.: Molecular Epidemiology of Bartonella vinsonii subsp. berkhoffii Infection in Coyotes from Central Coastal California"

    Article Title: Coyotes (Canis latrans) as the Reservoir for a Human Pathogenic Bartonella sp.: Molecular Epidemiology of Bartonella vinsonii subsp. berkhoffii Infection in Coyotes from Central Coastal California

    Journal: Journal of Clinical Microbiology

    doi:

    PCR-RFLP analysis of the 16S rRNA gene of coyote isolates with Dde I (A) or Mnl I (B) restriction endonuclease. Lanes 1 to 12, coyote isolates; lane 13, B. vinsonii subsp. berkhoffii ATCC 51672; lane 14, 100-bp molecular ladder.
    Figure Legend Snippet: PCR-RFLP analysis of the 16S rRNA gene of coyote isolates with Dde I (A) or Mnl I (B) restriction endonuclease. Lanes 1 to 12, coyote isolates; lane 13, B. vinsonii subsp. berkhoffii ATCC 51672; lane 14, 100-bp molecular ladder.

    Techniques Used: Polymerase Chain Reaction

    3) Product Images from "Clearance of an immunosuppressive virus from the CNS coincides with immune reanimation and diversification"

    Article Title: Clearance of an immunosuppressive virus from the CNS coincides with immune reanimation and diversification

    Journal: Virology Journal

    doi: 10.1186/1743-422X-4-53

    Neurotropic LCMV Armstrong is not selected for in the CNS of clone 13 infected mice . RNA was isolated from LCMV clones extracted from the brains of mice at 21 (n = 7 clones) and 150 days (n = 6 clones) post-clone 13 infection. RT-PCR, PCR and Mnl I restriction enzyme digests were performed as described in the Materials and Methods. The RNA PCR product from the Armstrong GP contains a phenylalanine at position 260 and is not cleaved by Mnl I. In contrast, clone 13 contains a leucine at position 260, and the 362 bp PCR product is cleaved into fragments (202 and 160 bp) by Mnl I. Note that all clones analyzed at both time points had the Mnl I restriction enzyme site. The control lane shows undigested 362 bp GP PCR product.
    Figure Legend Snippet: Neurotropic LCMV Armstrong is not selected for in the CNS of clone 13 infected mice . RNA was isolated from LCMV clones extracted from the brains of mice at 21 (n = 7 clones) and 150 days (n = 6 clones) post-clone 13 infection. RT-PCR, PCR and Mnl I restriction enzyme digests were performed as described in the Materials and Methods. The RNA PCR product from the Armstrong GP contains a phenylalanine at position 260 and is not cleaved by Mnl I. In contrast, clone 13 contains a leucine at position 260, and the 362 bp PCR product is cleaved into fragments (202 and 160 bp) by Mnl I. Note that all clones analyzed at both time points had the Mnl I restriction enzyme site. The control lane shows undigested 362 bp GP PCR product.

    Techniques Used: Infection, Mouse Assay, Isolation, Clone Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    4) Product Images from "Telomere damage induces internal loops that generate telomeric circles"

    Article Title: Telomere damage induces internal loops that generate telomeric circles

    Journal: bioRxiv

    doi: 10.1101/2020.01.29.924951

    A two-step procedure for the purification of mammalian telomeres A. Top: agarose gel showing the separation of the large telomeric repeat fragments from the bulk DNA in a sucrose gradient. Genomic DNA (~2.5 mg) from SV40-MEFs was digested with HinfI and MspI. The digested DNA was separated by centrifugation on a sucrose gradient. Seven fractions were collected and an aliquot (~1/500) of each fraction was loaded on an agarose gel. Bottom: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the high molecular weight (HMW) fractions. B. Left: agarose gel showing the separation of the large telomeric repeat fragments from the remaining non-telomeric DNA, in the second purification round. The HMW DNA, contained in the last four fractions of the sucrose gradient described in (A), was recovered and digested with RsaI, AluI, MboI, HinfI, MspI, HphI and MnlI. The digested DNA was separated on a preparative agarose gel and the DNA migrating in the area above 5 kb was extracted from the gel. The image shows an aliquot (~1/100) of the digested DNA, separated on an agarose gel. Right: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the HMW area. C. Dot blot analysis showing the enrichment of telomeric repeats. The indicated amounts of DNA from each enrichment step were spotted on a membrane and hybridized either with a probe recognizing the long interspersed BamHI repeats or TTAGGG repeats. The amount of TTAGGG repeat signal/ng was quantified and reported relative to the signal/ng value in the initial, non-enriched DNA. D. Single molecule analysis showing the enrichment of the telomeric repeats. The DNA was combed onto silanized coverslips, denatured in situ and labeled sequentially with an antibody against single-stranded DNA and a Cy3-labeled (TTAGGG) 3 PNA probe.
    Figure Legend Snippet: A two-step procedure for the purification of mammalian telomeres A. Top: agarose gel showing the separation of the large telomeric repeat fragments from the bulk DNA in a sucrose gradient. Genomic DNA (~2.5 mg) from SV40-MEFs was digested with HinfI and MspI. The digested DNA was separated by centrifugation on a sucrose gradient. Seven fractions were collected and an aliquot (~1/500) of each fraction was loaded on an agarose gel. Bottom: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the high molecular weight (HMW) fractions. B. Left: agarose gel showing the separation of the large telomeric repeat fragments from the remaining non-telomeric DNA, in the second purification round. The HMW DNA, contained in the last four fractions of the sucrose gradient described in (A), was recovered and digested with RsaI, AluI, MboI, HinfI, MspI, HphI and MnlI. The digested DNA was separated on a preparative agarose gel and the DNA migrating in the area above 5 kb was extracted from the gel. The image shows an aliquot (~1/100) of the digested DNA, separated on an agarose gel. Right: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the HMW area. C. Dot blot analysis showing the enrichment of telomeric repeats. The indicated amounts of DNA from each enrichment step were spotted on a membrane and hybridized either with a probe recognizing the long interspersed BamHI repeats or TTAGGG repeats. The amount of TTAGGG repeat signal/ng was quantified and reported relative to the signal/ng value in the initial, non-enriched DNA. D. Single molecule analysis showing the enrichment of the telomeric repeats. The DNA was combed onto silanized coverslips, denatured in situ and labeled sequentially with an antibody against single-stranded DNA and a Cy3-labeled (TTAGGG) 3 PNA probe.

    Techniques Used: Purification, Agarose Gel Electrophoresis, Centrifugation, Molecular Weight, Dot Blot, In Situ, Labeling

    5) Product Images from "Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba"

    Article Title: Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.69.7.4159-4166.2003

    TRFLP profiles of Mnl I-digested 16S rDNAs amplified from different anaerobic cultures enriched on brominated phenolic compounds. (A) Native sponge-associated microorganisms; (B) control with only 200 μM lactate; (C) a mixture of 2-, 3- and 4-BP (100 μM each); (D) 100 μM 2-BP; (E) 100 μM 3-BP; (F) 100 μM 4-BP; (G) 100 μM 3,5-DB-4-HB; (H) 100 μM 2,6-DBP; and (I) 100 μM 2,4,6-TBP.
    Figure Legend Snippet: TRFLP profiles of Mnl I-digested 16S rDNAs amplified from different anaerobic cultures enriched on brominated phenolic compounds. (A) Native sponge-associated microorganisms; (B) control with only 200 μM lactate; (C) a mixture of 2-, 3- and 4-BP (100 μM each); (D) 100 μM 2-BP; (E) 100 μM 3-BP; (F) 100 μM 4-BP; (G) 100 μM 3,5-DB-4-HB; (H) 100 μM 2,6-DBP; and (I) 100 μM 2,4,6-TBP.

    Techniques Used: Terminal Restriction Fragment Length Polymorphism, Amplification

    6) Product Images from "Targeting Single-Nucleotide Polymorphisms in the 18S rRNA Gene To Differentiate Cyclospora Species from Eimeria Species by Multiplex PCR"

    Article Title: Targeting Single-Nucleotide Polymorphisms in the 18S rRNA Gene To Differentiate Cyclospora Species from Eimeria Species by Multiplex PCR

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.69.8.4806-4813.2003

    Molecular identification of Cyclospora and Eimeria species. (A) Conventional nested-PCR amplification. Partially purified oocysts (100 to 1,000) were spotted onto FTA filters that were used as a PCR template. For the primate-derived Cyclospora -like organisms, 2 μl of purified DNA was used as a template. A primary amplicon (636 bp [not shown]) was generated from each of these templates with the F1E-R2B primer pair, and 1 μl of this product was used in a nested amplification with the F3E-R4B primer pair to generate the 294-bp amplicon shown. (B) RFLP analysis of nested-PCR amplicons. Nested-PCR amplicons were digested with Mnl I and analyzed by gel electrophoresis with 5% NuSieve 3:1 agarose containing ethidium bromide (0.2 μg/ml).
    Figure Legend Snippet: Molecular identification of Cyclospora and Eimeria species. (A) Conventional nested-PCR amplification. Partially purified oocysts (100 to 1,000) were spotted onto FTA filters that were used as a PCR template. For the primate-derived Cyclospora -like organisms, 2 μl of purified DNA was used as a template. A primary amplicon (636 bp [not shown]) was generated from each of these templates with the F1E-R2B primer pair, and 1 μl of this product was used in a nested amplification with the F3E-R4B primer pair to generate the 294-bp amplicon shown. (B) RFLP analysis of nested-PCR amplicons. Nested-PCR amplicons were digested with Mnl I and analyzed by gel electrophoresis with 5% NuSieve 3:1 agarose containing ethidium bromide (0.2 μg/ml).

    Techniques Used: Nested PCR, Amplification, Purification, Polymerase Chain Reaction, Derivative Assay, Generated, Nucleic Acid Electrophoresis

    Analysis of a human fecal specimen by multiplex PCR amplification by using SNP primers to diagnose a C. cayetanensis infection. Raw fecal material (10 μl) was spotted onto an FTA filter and prepared for PCR. A primary amplicon was generated with the F1E-R2B primer pair by conventional PCR and then used in subsequent nested amplifications. (A) Microscopic identification (1000x) of C. cayetanensis by using Differential interference contrast (left panel) and UV autofluorescence (right panel) images. Magnification, ×800. (B) Multiplex PCR amplification with SNP primers. Lane a, 100-bp DNA ladder; lane b, SNP multiplex PCR amplicon standards from C. cercopitheci (360 bp), C. cayetanensis (300 bp), and E. tenella (173 bp); lane c, patient specimen. (C) Conventional nested-PCR amplification. Lanes a and f, 100-bp DNA ladder; lane b, C. cayetanensis ; lane c, C. cercopitheci ; lane d, E. tenella ; lane e, patient specimen. (D) RFLP analysis of nested-PCR amplicons. Nested amplicons obtained in panel B were digested with Mnl I. Lanes a and f, 20-bp DNA ladders; lane b, C. cayetanensis ; lane c, C. cercopitheci ; lane d, E. tenella ; lane e, patient specimen.
    Figure Legend Snippet: Analysis of a human fecal specimen by multiplex PCR amplification by using SNP primers to diagnose a C. cayetanensis infection. Raw fecal material (10 μl) was spotted onto an FTA filter and prepared for PCR. A primary amplicon was generated with the F1E-R2B primer pair by conventional PCR and then used in subsequent nested amplifications. (A) Microscopic identification (1000x) of C. cayetanensis by using Differential interference contrast (left panel) and UV autofluorescence (right panel) images. Magnification, ×800. (B) Multiplex PCR amplification with SNP primers. Lane a, 100-bp DNA ladder; lane b, SNP multiplex PCR amplicon standards from C. cercopitheci (360 bp), C. cayetanensis (300 bp), and E. tenella (173 bp); lane c, patient specimen. (C) Conventional nested-PCR amplification. Lanes a and f, 100-bp DNA ladder; lane b, C. cayetanensis ; lane c, C. cercopitheci ; lane d, E. tenella ; lane e, patient specimen. (D) RFLP analysis of nested-PCR amplicons. Nested amplicons obtained in panel B were digested with Mnl I. Lanes a and f, 20-bp DNA ladders; lane b, C. cayetanensis ; lane c, C. cercopitheci ; lane d, E. tenella ; lane e, patient specimen.

    Techniques Used: Multiplex Assay, Polymerase Chain Reaction, Amplification, Infection, Generated, Nested PCR

    7) Product Images from "Development and Evaluation of a PCR-Based Assay for Detection of Haemobartonella felis in Cats and Differentiation of H. felis from Related Bacteria by Restriction Fragment Length Polymorphism Analysis"

    Article Title: Development and Evaluation of a PCR-Based Assay for Detection of Haemobartonella felis in Cats and Differentiation of H. felis from Related Bacteria by Restriction Fragment Length Polymorphism Analysis

    Journal: Journal of Clinical Microbiology

    doi:

    Restriction profiles obtained after Dde I and Mnl I digestion of the 16S rRNA gene amplified with the universal primer set fHf1 and rHf1. Lanes: 1 to 4, Dde I digests of H. felis , B. bacilliformis , M. genitalium , and E. suis , respectively; 5 to 8, Mnl I digests of H. felis , B. bacilliformis , M. genitalium , and E. suis , respectively; M 1 and M 2 , molecular size markers (base pair values are indicated in the left and right margins).
    Figure Legend Snippet: Restriction profiles obtained after Dde I and Mnl I digestion of the 16S rRNA gene amplified with the universal primer set fHf1 and rHf1. Lanes: 1 to 4, Dde I digests of H. felis , B. bacilliformis , M. genitalium , and E. suis , respectively; 5 to 8, Mnl I digests of H. felis , B. bacilliformis , M. genitalium , and E. suis , respectively; M 1 and M 2 , molecular size markers (base pair values are indicated in the left and right margins).

    Techniques Used: Amplification

    Related Articles

    Amplification:

    Article Title: Coyotes (Canis latrans) as the Reservoir for a Human Pathogenic Bartonella sp.: Molecular Epidemiology of Bartonella vinsonii subsp. berkhoffii Infection in Coyotes from Central Coastal California
    Article Snippet: .. The amplified product of the 16S rRNA gene was digested with Dde I (Boehringer GmbH, Mannheim, Germany) and Mnl I (New England BioLabs) restriction endonucleases. .. Banding patterns were compared with those of a domestic dog isolate (American Type Culture Collection [ATCC] 51672) of B. vinsonii subsp. berkhoffii ), B. vinsonii subsp. vinsonii (ATCC VR152), B. henselae (strain U-4; University of California, Davis) and B. clarridgeiae (ATCC 51734); the last two Bartonella species are usually isolated from domestic cats.

    Agarose Gel Electrophoresis:

    Article Title: Clearance of an immunosuppressive virus from the CNS coincides with immune reanimation and diversification
    Article Snippet: .. 10 μg of the PCR product were digested with Mnl I (NEB) and analyzed by agarose gel electrophoresis. ..

    Article Title: Telomere damage induces internal loops that generate telomeric circles
    Article Snippet: .. The DNA was then digested overnight with 50 units each of RsaI, AluI, MboI, HinfI, MspI, HphI, MnlI (NEB) and then separated on a 0.7% low-melting agarose gel (SeaPlaque Agarose, Lonza, 50100), without ethidium bromide. .. Fragments migrating above the 5 kb band of the marker were extracted using the Silica Bead DNA gel extraction kit (Thermo Fisher Scientific, K0513) following the manufacturer’s instructions, except that once the DNA was bound, the beads were not resuspended to avoid mechanical shearing of the DNA.

    Generated:

    Article Title: Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings
    Article Snippet: .. For this assay, 10 µL genomic PCR products were generated with the RNF168 primer pair listed above and were incubated overnight with 1.5 U Mnl I (New England Biolabs). .. Cleavage fragments were separated through electrophoresis on 2% agarose gels supplemented with GelRed and were visualized over an UV transilluminator.

    Labeling:

    Article Title: Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba
    Article Snippet: .. Fluorescently labeled PCR products (20 μl) were purified with the Geneclean kit II (Qbiogene, Inc., Carlsbad, Calif.) and were digested for 6 h at 37°C with Mnl I (New England Biolabs, Beverly, Mass.). .. Twenty-five nanograms of labeled PCR product was run on a ABI 310 genetic analyzer (Perkin-Elmer) with Genescan software and internal standards ( , ).

    Purification:

    Article Title: Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba
    Article Snippet: .. Fluorescently labeled PCR products (20 μl) were purified with the Geneclean kit II (Qbiogene, Inc., Carlsbad, Calif.) and were digested for 6 h at 37°C with Mnl I (New England Biolabs, Beverly, Mass.). .. Twenty-five nanograms of labeled PCR product was run on a ABI 310 genetic analyzer (Perkin-Elmer) with Genescan software and internal standards ( , ).

    Incubation:

    Article Title: Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings
    Article Snippet: .. For this assay, 10 µL genomic PCR products were generated with the RNF168 primer pair listed above and were incubated overnight with 1.5 U Mnl I (New England Biolabs). .. Cleavage fragments were separated through electrophoresis on 2% agarose gels supplemented with GelRed and were visualized over an UV transilluminator.

    Polymerase Chain Reaction:

    Article Title: Clearance of an immunosuppressive virus from the CNS coincides with immune reanimation and diversification
    Article Snippet: .. 10 μg of the PCR product were digested with Mnl I (NEB) and analyzed by agarose gel electrophoresis. ..

    Article Title: Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings
    Article Snippet: .. For this assay, 10 µL genomic PCR products were generated with the RNF168 primer pair listed above and were incubated overnight with 1.5 U Mnl I (New England Biolabs). .. Cleavage fragments were separated through electrophoresis on 2% agarose gels supplemented with GelRed and were visualized over an UV transilluminator.

    Article Title: Reductive Dehalogenation of Brominated Phenolic Compounds by Microorganisms Associated with the Marine Sponge Aplysina aerophoba
    Article Snippet: .. Fluorescently labeled PCR products (20 μl) were purified with the Geneclean kit II (Qbiogene, Inc., Carlsbad, Calif.) and were digested for 6 h at 37°C with Mnl I (New England Biolabs, Beverly, Mass.). .. Twenty-five nanograms of labeled PCR product was run on a ABI 310 genetic analyzer (Perkin-Elmer) with Genescan software and internal standards ( , ).

    Article Title: Identification of Unique Type II Polyketide Synthase Genes in Soil
    Article Snippet: .. FAM-labeled PCR products were diluted to 5 ng μl−1 and 30 ng were independently digested for 4 h using 2 units of MnlI or AluI (NEB, Beverly, MA). ..

    Article Title: Development and Evaluation of a PCR-Based Assay for Detection of Haemobartonella felis in Cats and Differentiation of H. felis from Related Bacteria by Restriction Fragment Length Polymorphism Analysis
    Article Snippet: .. Two restriction enzymes, Dde I and Mnl I (New England Biolabs, Beverly, Mass.), were used according to the manufacturer’s recommendations to digest the PCR products. .. Dde I recognizes the sequence 5′-C▾ TNAG-3′ and cleaves at the position indicated by the arrowhead ( ).

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  • 98
    New England Biolabs mnl i
    Assessment of <t>RNF168</t> mutation (A) and RNF168 protein level (B) . (A) Direct sequencing of RNF168 exon 12 reveals homozygosity for the novel frameshift mutation c.295delG in genomic DNA from either of both patients (HA591, HA592). (B) RFLP analysis of PCR products on 2% agarose gel electrophoresis. S, size marker; 1, undigested PCR product; 2–6, PCR products cleaved with Mnl I: 2, wild-type control, 3, paternal sample, 4. maternal sample, 5, patient HA591, 6, patient HA592. (C) Western blot analysis reveals strongly reduced immunoreactivity for RNF168 protein in lymphoblastoid cells from either of both patients (HA591, HA592). Lymphoblastoid cell lines (LCLs) from a healthy individual were used as an RNF168-proficient control (HA325), and LCLs from a patient with classical ataxia–telangiectasia were used for comparison (HA56). β-actin served as the loading control (ACTB).
    Mnl I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mnl i/product/New England Biolabs
    Average 98 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    mnl i - by Bioz Stars, 2020-09
    98/100 stars
      Buy from Supplier

    Image Search Results


    Assessment of RNF168 mutation (A) and RNF168 protein level (B) . (A) Direct sequencing of RNF168 exon 12 reveals homozygosity for the novel frameshift mutation c.295delG in genomic DNA from either of both patients (HA591, HA592). (B) RFLP analysis of PCR products on 2% agarose gel electrophoresis. S, size marker; 1, undigested PCR product; 2–6, PCR products cleaved with Mnl I: 2, wild-type control, 3, paternal sample, 4. maternal sample, 5, patient HA591, 6, patient HA592. (C) Western blot analysis reveals strongly reduced immunoreactivity for RNF168 protein in lymphoblastoid cells from either of both patients (HA591, HA592). Lymphoblastoid cell lines (LCLs) from a healthy individual were used as an RNF168-proficient control (HA325), and LCLs from a patient with classical ataxia–telangiectasia were used for comparison (HA56). β-actin served as the loading control (ACTB).

    Journal: Frontiers in Immunology

    Article Title: Clinical and Biological Manifestation of RNF168 Deficiency in Two Polish Siblings

    doi: 10.3389/fimmu.2017.01683

    Figure Lengend Snippet: Assessment of RNF168 mutation (A) and RNF168 protein level (B) . (A) Direct sequencing of RNF168 exon 12 reveals homozygosity for the novel frameshift mutation c.295delG in genomic DNA from either of both patients (HA591, HA592). (B) RFLP analysis of PCR products on 2% agarose gel electrophoresis. S, size marker; 1, undigested PCR product; 2–6, PCR products cleaved with Mnl I: 2, wild-type control, 3, paternal sample, 4. maternal sample, 5, patient HA591, 6, patient HA592. (C) Western blot analysis reveals strongly reduced immunoreactivity for RNF168 protein in lymphoblastoid cells from either of both patients (HA591, HA592). Lymphoblastoid cell lines (LCLs) from a healthy individual were used as an RNF168-proficient control (HA325), and LCLs from a patient with classical ataxia–telangiectasia were used for comparison (HA56). β-actin served as the loading control (ACTB).

    Article Snippet: For this assay, 10 µL genomic PCR products were generated with the RNF168 primer pair listed above and were incubated overnight with 1.5 U Mnl I (New England Biolabs).

    Techniques: Mutagenesis, Sequencing, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker, Western Blot

    PCR-RFLP analysis of the 16S rRNA gene of coyote isolates with Dde I (A) or Mnl I (B) restriction endonuclease. Lanes 1 to 12, coyote isolates; lane 13, B. vinsonii subsp. berkhoffii ATCC 51672; lane 14, 100-bp molecular ladder.

    Journal: Journal of Clinical Microbiology

    Article Title: Coyotes (Canis latrans) as the Reservoir for a Human Pathogenic Bartonella sp.: Molecular Epidemiology of Bartonella vinsonii subsp. berkhoffii Infection in Coyotes from Central Coastal California

    doi:

    Figure Lengend Snippet: PCR-RFLP analysis of the 16S rRNA gene of coyote isolates with Dde I (A) or Mnl I (B) restriction endonuclease. Lanes 1 to 12, coyote isolates; lane 13, B. vinsonii subsp. berkhoffii ATCC 51672; lane 14, 100-bp molecular ladder.

    Article Snippet: The amplified product of the 16S rRNA gene was digested with Dde I (Boehringer GmbH, Mannheim, Germany) and Mnl I (New England BioLabs) restriction endonucleases.

    Techniques: Polymerase Chain Reaction

    Neurotropic LCMV Armstrong is not selected for in the CNS of clone 13 infected mice . RNA was isolated from LCMV clones extracted from the brains of mice at 21 (n = 7 clones) and 150 days (n = 6 clones) post-clone 13 infection. RT-PCR, PCR and Mnl I restriction enzyme digests were performed as described in the Materials and Methods. The RNA PCR product from the Armstrong GP contains a phenylalanine at position 260 and is not cleaved by Mnl I. In contrast, clone 13 contains a leucine at position 260, and the 362 bp PCR product is cleaved into fragments (202 and 160 bp) by Mnl I. Note that all clones analyzed at both time points had the Mnl I restriction enzyme site. The control lane shows undigested 362 bp GP PCR product.

    Journal: Virology Journal

    Article Title: Clearance of an immunosuppressive virus from the CNS coincides with immune reanimation and diversification

    doi: 10.1186/1743-422X-4-53

    Figure Lengend Snippet: Neurotropic LCMV Armstrong is not selected for in the CNS of clone 13 infected mice . RNA was isolated from LCMV clones extracted from the brains of mice at 21 (n = 7 clones) and 150 days (n = 6 clones) post-clone 13 infection. RT-PCR, PCR and Mnl I restriction enzyme digests were performed as described in the Materials and Methods. The RNA PCR product from the Armstrong GP contains a phenylalanine at position 260 and is not cleaved by Mnl I. In contrast, clone 13 contains a leucine at position 260, and the 362 bp PCR product is cleaved into fragments (202 and 160 bp) by Mnl I. Note that all clones analyzed at both time points had the Mnl I restriction enzyme site. The control lane shows undigested 362 bp GP PCR product.

    Article Snippet: 10 μg of the PCR product were digested with Mnl I (NEB) and analyzed by agarose gel electrophoresis.

    Techniques: Infection, Mouse Assay, Isolation, Clone Assay, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    A two-step procedure for the purification of mammalian telomeres A. Top: agarose gel showing the separation of the large telomeric repeat fragments from the bulk DNA in a sucrose gradient. Genomic DNA (~2.5 mg) from SV40-MEFs was digested with HinfI and MspI. The digested DNA was separated by centrifugation on a sucrose gradient. Seven fractions were collected and an aliquot (~1/500) of each fraction was loaded on an agarose gel. Bottom: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the high molecular weight (HMW) fractions. B. Left: agarose gel showing the separation of the large telomeric repeat fragments from the remaining non-telomeric DNA, in the second purification round. The HMW DNA, contained in the last four fractions of the sucrose gradient described in (A), was recovered and digested with RsaI, AluI, MboI, HinfI, MspI, HphI and MnlI. The digested DNA was separated on a preparative agarose gel and the DNA migrating in the area above 5 kb was extracted from the gel. The image shows an aliquot (~1/100) of the digested DNA, separated on an agarose gel. Right: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the HMW area. C. Dot blot analysis showing the enrichment of telomeric repeats. The indicated amounts of DNA from each enrichment step were spotted on a membrane and hybridized either with a probe recognizing the long interspersed BamHI repeats or TTAGGG repeats. The amount of TTAGGG repeat signal/ng was quantified and reported relative to the signal/ng value in the initial, non-enriched DNA. D. Single molecule analysis showing the enrichment of the telomeric repeats. The DNA was combed onto silanized coverslips, denatured in situ and labeled sequentially with an antibody against single-stranded DNA and a Cy3-labeled (TTAGGG) 3 PNA probe.

    Journal: bioRxiv

    Article Title: Telomere damage induces internal loops that generate telomeric circles

    doi: 10.1101/2020.01.29.924951

    Figure Lengend Snippet: A two-step procedure for the purification of mammalian telomeres A. Top: agarose gel showing the separation of the large telomeric repeat fragments from the bulk DNA in a sucrose gradient. Genomic DNA (~2.5 mg) from SV40-MEFs was digested with HinfI and MspI. The digested DNA was separated by centrifugation on a sucrose gradient. Seven fractions were collected and an aliquot (~1/500) of each fraction was loaded on an agarose gel. Bottom: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the high molecular weight (HMW) fractions. B. Left: agarose gel showing the separation of the large telomeric repeat fragments from the remaining non-telomeric DNA, in the second purification round. The HMW DNA, contained in the last four fractions of the sucrose gradient described in (A), was recovered and digested with RsaI, AluI, MboI, HinfI, MspI, HphI and MnlI. The digested DNA was separated on a preparative agarose gel and the DNA migrating in the area above 5 kb was extracted from the gel. The image shows an aliquot (~1/100) of the digested DNA, separated on an agarose gel. Right: the gel was blotted onto a membrane and hybridized with a TTAGGG repeats probe to verify that telomeric repeats remained in the HMW area. C. Dot blot analysis showing the enrichment of telomeric repeats. The indicated amounts of DNA from each enrichment step were spotted on a membrane and hybridized either with a probe recognizing the long interspersed BamHI repeats or TTAGGG repeats. The amount of TTAGGG repeat signal/ng was quantified and reported relative to the signal/ng value in the initial, non-enriched DNA. D. Single molecule analysis showing the enrichment of the telomeric repeats. The DNA was combed onto silanized coverslips, denatured in situ and labeled sequentially with an antibody against single-stranded DNA and a Cy3-labeled (TTAGGG) 3 PNA probe.

    Article Snippet: The DNA was then digested overnight with 50 units each of RsaI, AluI, MboI, HinfI, MspI, HphI, MnlI (NEB) and then separated on a 0.7% low-melting agarose gel (SeaPlaque Agarose, Lonza, 50100), without ethidium bromide.

    Techniques: Purification, Agarose Gel Electrophoresis, Centrifugation, Molecular Weight, Dot Blot, In Situ, Labeling