cviki 1  (New England Biolabs)


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  • 94
    Name:
    CviKI 1
    Description:
    CviKI 1 250 units
    Catalog Number:
    r0710l
    Price:
    70
    Size:
    250 units
    Category:
    Restriction Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs cviki 1
    CviKI 1
    CviKI 1 250 units
    https://www.bioz.com/result/cviki 1/product/New England Biolabs
    Average 94 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    cviki 1 - by Bioz Stars, 2020-07
    94/100 stars

    Images

    1) Product Images from "RELACS nuclei barcoding enables high-throughput ChIP-seq"

    Article Title: RELACS nuclei barcoding enables high-throughput ChIP-seq

    Journal: Communications Biology

    doi: 10.1038/s42003-018-0219-z

    RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days
    Figure Legend Snippet: RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days

    Techniques Used: Sonication, Concentration Assay, Incubation, Chromatin Immunoprecipitation, DNA Purification, Polymerase Chain Reaction, Amplification, Generated

    2) Product Images from "Characterization of the Chromosome 1q41q42.12 region, and the Candidate Gene DISP1, in Patients with CDH"

    Article Title: Characterization of the Chromosome 1q41q42.12 region, and the Candidate Gene DISP1, in Patients with CDH

    Journal: American journal of medical genetics. Part A

    doi: 10.1002/ajmg.a.33618

    Restriction enzyme digestion of a 195-bp DISP1 exon 8 product with CviKI-1. Wild type product with CviKI-1 recognition site (GG ↓CT) is digested into fragments of 103-bp and 92-bp. The mutation [c.4412C > G (p.Ala1471Gly)] abolishes a CviKI-1
    Figure Legend Snippet: Restriction enzyme digestion of a 195-bp DISP1 exon 8 product with CviKI-1. Wild type product with CviKI-1 recognition site (GG ↓CT) is digested into fragments of 103-bp and 92-bp. The mutation [c.4412C > G (p.Ala1471Gly)] abolishes a CviKI-1

    Techniques Used: Mutagenesis

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Characterization of the Chromosome 1q41q42.12 region, and the Candidate Gene DISP1, in Patients with CDH
    Article Snippet: .. The PCR products using forward primer 5′-GGAAAGTGGAGCTGAGCTTG-3′ and reverse primer 5′-CCATGTTTGGCATTTGACAG-3′ were digested by the restriction endonuclease, CviKI-1 enzyme (New England Biolabs, Ipswich, MA) which specifically recognizes and cleaves the nucleotide sequence 5′-RG↓CY-3′. .. The 10μl of PCR products (195bp) were incubated with CviKI-1 (10U) overnight at 37°C.

    Article Title: The -258 A/G (SNP rs12885300) polymorphism of the human type-2 deiodinase gene is associated with a shift in the pattern of secretion of thyroid hormones following a TRH-induced acute rise in TSH
    Article Snippet: .. After purification, PCR products were digested with CviKI-1 (New England BioLabs, Ipswich, MA), restriction enzyme for 4 hours at 37°C. .. The digestion products were separated on 4.5% agarose gel by electrophoresis and visualized by ethidium bromide staining.

    Article Title: Complete chloroplast genome sequence and phylogenetic analysis of wasabi (Eutrema japonicum) and its relatives
    Article Snippet: .. The PCR products were digested at two or three sites by the restriction enzyme, CviKI -1 (New England Biolabs, Ipswich, MA). .. As expected from the cp nucleotide sequences, three bands (227, 386, and 407 bp) were generated in ‘Fujidaruma’ and ‘Shimane No. 3’, whereas three bands of 227, 350 and 386 bp in size were detected in the other cultivated accessions (Fig. ).

    Sequencing:

    Article Title: Characterization of the Chromosome 1q41q42.12 region, and the Candidate Gene DISP1, in Patients with CDH
    Article Snippet: .. The PCR products using forward primer 5′-GGAAAGTGGAGCTGAGCTTG-3′ and reverse primer 5′-CCATGTTTGGCATTTGACAG-3′ were digested by the restriction endonuclease, CviKI-1 enzyme (New England Biolabs, Ipswich, MA) which specifically recognizes and cleaves the nucleotide sequence 5′-RG↓CY-3′. .. The 10μl of PCR products (195bp) were incubated with CviKI-1 (10U) overnight at 37°C.

    Incubation:

    Article Title: SMASH, a fragmentation and sequencing method for genomic copy number analysis
    Article Snippet: .. DNA fragments were further cut by CviKI-1 (NEB) and NlaIII (NEB) in 1× CutSmart buffer in a final volume of 90 µL, which was incubated at 37°C for 1 h. After enzymatic digestion, the solution volume was reduced to ∼30 µL by Savant SpeedVac (Thermo Scientific). .. DNA fragments > 100 bp were removed as follows: adding 2.5× volume of AMPure XP beads (Beckman Coulter), mixing well, incubating at room temperature (RT) for 5 min, and collecting the supernatant.

    Clone Assay:

    Article Title: Application of M13 phage display for identifying immunogenic proteins from tick (Ixodes scapularis) saliva
    Article Snippet: .. Salivary gland library construction To generate blunt ended DNA fragments suitable for cloning into pHORF3 [ ] a restriction enzyme digest using the blunt end cutting enzymes AfeI, AluI and CviKI-1 (NEB, Frankfurt, Germany) was performed. .. 30 μl SMART cDNA (~1.7 μg) were mixed with 1.25 U of each of the restriction enzymes, 10 μL NEBuffer 4 (NEB), 10 μL 10xBSA solution (NEB) in a total volume of 100 μl.

    Purification:

    Article Title: The -258 A/G (SNP rs12885300) polymorphism of the human type-2 deiodinase gene is associated with a shift in the pattern of secretion of thyroid hormones following a TRH-induced acute rise in TSH
    Article Snippet: .. After purification, PCR products were digested with CviKI-1 (New England BioLabs, Ipswich, MA), restriction enzyme for 4 hours at 37°C. .. The digestion products were separated on 4.5% agarose gel by electrophoresis and visualized by ethidium bromide staining.

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    New England Biolabs cviki 1
    RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work <t>CviKI-1</t> was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days
    Cviki 1, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cviki 1/product/New England Biolabs
    Average 94 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    cviki 1 - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

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    RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days

    Journal: Communications Biology

    Article Title: RELACS nuclei barcoding enables high-throughput ChIP-seq

    doi: 10.1038/s42003-018-0219-z

    Figure Lengend Snippet: RELACS workflow. Overview of the RELACS method. The protocol facilitates barcoding multiple cell populations, which can be pooled and investigated for multiple epitopes within the same run. The method starts by isolating nuclei from a pool of formaldehyde-fixed cells, using sonication to reduce cell type dependency 5 ( a ). The nuclear membrane is permeabilized to allow entrance of enzymes, followed by DNA barcodes. Restriction endonucleases with a high frequency of recognition sites are used to fragment chromatin—in this work CviKI-1 was used ( b ). Nuclei are washed to remove active restriction enzymes ( c ). Hairpin adapters harboring barcodes are ligated to both the ends of the fragmented chromatin inside the nuclei. The barcoding has been tested using 100–500,000 nuclei without the need to change protocol conditions ( d ). Cell populations marked with specific barcodes are pooled ( e ), concentrated and lysed using SDS at low concentration and short sonication to release chromatin into solution ( f ). Chromatin is split and incubated with the antibodies of interest ( g ). After ChIP washes and DNA purification (not illustrated), only DNA that harbors nuclei barcodes at both ends is PCR amplified to complete library construction. PCR amplification appends an Illumina barcode onto mark each fragment ( h ). Sequenced libraries are demultiplexed by Illumina barcode, to retrieve ChIP information, and then nuclear barcode, to identify the initial cell population ( i ). The RELACS protocol is very fast and ChIP-seq libraries can be generated for hundreds of samples within three days

    Article Snippet: Nuclei digestion Five units of the restriction enzyme CviKI-1 (NEB, R0710S) were added to every 100,000 nuclei.

    Techniques: Sonication, Concentration Assay, Incubation, Chromatin Immunoprecipitation, DNA Purification, Polymerase Chain Reaction, Amplification, Generated

    Restriction enzyme digestion of a 195-bp DISP1 exon 8 product with CviKI-1. Wild type product with CviKI-1 recognition site (GG ↓CT) is digested into fragments of 103-bp and 92-bp. The mutation [c.4412C > G (p.Ala1471Gly)] abolishes a CviKI-1

    Journal: American journal of medical genetics. Part A

    Article Title: Characterization of the Chromosome 1q41q42.12 region, and the Candidate Gene DISP1, in Patients with CDH

    doi: 10.1002/ajmg.a.33618

    Figure Lengend Snippet: Restriction enzyme digestion of a 195-bp DISP1 exon 8 product with CviKI-1. Wild type product with CviKI-1 recognition site (GG ↓CT) is digested into fragments of 103-bp and 92-bp. The mutation [c.4412C > G (p.Ala1471Gly)] abolishes a CviKI-1

    Article Snippet: The PCR products using forward primer 5′-GGAAAGTGGAGCTGAGCTTG-3′ and reverse primer 5′-CCATGTTTGGCATTTGACAG-3′ were digested by the restriction endonuclease, CviKI-1 enzyme (New England Biolabs, Ipswich, MA) which specifically recognizes and cleaves the nucleotide sequence 5′-RG↓CY-3′.

    Techniques: Mutagenesis