kpni  (New England Biolabs)


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  • 99
    Name:
    KpnI
    Description:
    KpnI 20 000 units
    Catalog Number:
    r0142l
    Price:
    264
    Size:
    20 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs kpni
    KpnI
    KpnI 20 000 units
    https://www.bioz.com/result/kpni/product/New England Biolabs
    Average 99 stars, based on 233 article reviews
    Price from $9.99 to $1999.99
    kpni - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Transduction:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. Lentiviruses made from the sublibrary were then used to transduce into CHOP-mCherry reporter cells, which were subjected to a new round of As treatment and sorting.

    Clone Assay:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625. .. A 15-nt 5′-extension (GGC CTT CGG GCC AAG) was added to these gRNA constructs to facilitate SHAPE analysis ( ).

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: .. Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. Similarly, to generate pcDNA 3.1-puro-mCherry, PCR amplified mCherry was cloned into EcoRI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    Amplification:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: For PCR amplification the following conditions were used: initial denaturation at 98 °C for 4 min, 98 °C for 20 s, 30 cycles at 60 °C for 20 s, 72 °C for 30 s, and final extension at 72 °C for 3 min. .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies).

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: .. Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. Similarly, to generate pcDNA 3.1-puro-mCherry, PCR amplified mCherry was cloned into EcoRI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    Construct:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625. .. A 15-nt 5′-extension (GGC CTT CGG GCC AAG) was added to these gRNA constructs to facilitate SHAPE analysis ( ).

    Article Title: Next Generation of Tn7-Based Single-Copy Insertion Elements for Use in Multi- and Pan-Drug-Resistant Strains of Acinetobacter baumannii
    Article Snippet: The genes for the fluorescent proteins along with their native promoters were excised from their parent plasmids ( ) using HindIII and KpnI (New England Biolabs) and ligated to pUC18T-mini-Tn 7 T-Apr digested with the same enzymes before transformation into E. coli DH5α. .. The constructs were confirmed via restriction digestion, and maps of these plasmids are provided in Fig. S2 in the supplemental material.

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: .. Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. Similarly, to generate pcDNA 3.1-puro-mCherry, PCR amplified mCherry was cloned into EcoRI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    Transformation Assay:

    Article Title: Next Generation of Tn7-Based Single-Copy Insertion Elements for Use in Multi- and Pan-Drug-Resistant Strains of Acinetobacter baumannii
    Article Snippet: .. The genes for the fluorescent proteins along with their native promoters were excised from their parent plasmids ( ) using HindIII and KpnI (New England Biolabs) and ligated to pUC18T-mini-Tn 7 T-Apr digested with the same enzymes before transformation into E. coli DH5α. ..

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. Lentiviruses made from the sublibrary were then used to transduce into CHOP-mCherry reporter cells, which were subjected to a new round of As treatment and sorting.

    Derivative Assay:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625. .. A 15-nt 5′-extension (GGC CTT CGG GCC AAG) was added to these gRNA constructs to facilitate SHAPE analysis ( ).

    Countercurrent Chromatography:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: HTLV-1 SL(450–470) (5′-AUG GGC CAA AUC UUU UCC CGU UU -3′) and SL(474–508) (5′-GCU AGC CCU AUU CCG CGG CCG CCC CGG GGG CUG GC U U -3′) were purchased from Dharmacon (GE healthcare). .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625.

    Generated:

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. GFP-tagged pcDNA5/FRT and pcDNA5/FRT TO vector, having hygromycin as selectable marker, was generated by PCR sub-cloning of GFP into EcoRI and EcoRV (New England Biolabs) site of pcDNA5/FRT, Xho-1 site of pcDNA5/FRT TO vector respectively.

    Polymerase Chain Reaction:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. Lentiviruses made from the sublibrary were then used to transduce into CHOP-mCherry reporter cells, which were subjected to a new round of As treatment and sorting.

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: .. Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. Similarly, to generate pcDNA 3.1-puro-mCherry, PCR amplified mCherry was cloned into EcoRI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    Cellular Antioxidant Activity Assay:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: HTLV-1 SL(450–470) (5′-AUG GGC CAA AUC UUU UCC CGU UU -3′) and SL(474–508) (5′-GCU AGC CCU AUU CCG CGG CCG CCC CGG GGG CUG GC U U -3′) were purchased from Dharmacon (GE healthcare). .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625.

    Fluorescence:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: The upper 10% (fluorescence intensity) of the mCherry-positive cell population was isolated, recovered, and allowed to repopulate for 4 d. The sorted reporter cells were treated again with 5 µM As and the upper 10% of the bright population was isolated. .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies).

    Mutagenesis:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625. .. To probe potential dimerization sites in the 5′ leader, three palindromic loop regions, nt 348–350, nt 382–386, and nt 498–500, were mutated to GAGA tetraloops individually and simultaneously using site-directed, ligase-independent mutagenesis (SLIM) ( ).

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: Paragraph title: Cloning and mutagenesis ... Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    Isolation:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: The upper 10% (fluorescence intensity) of the mCherry-positive cell population was isolated, recovered, and allowed to repopulate for 4 d. The sorted reporter cells were treated again with 5 µM As and the upper 10% of the bright population was isolated. .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies).

    Subcloning:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. Lentiviruses made from the sublibrary were then used to transduce into CHOP-mCherry reporter cells, which were subjected to a new round of As treatment and sorting.

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. GFP-tagged pcDNA5/FRT and pcDNA5/FRT TO vector, having hygromycin as selectable marker, was generated by PCR sub-cloning of GFP into EcoRI and EcoRV (New England Biolabs) site of pcDNA5/FRT, Xho-1 site of pcDNA5/FRT TO vector respectively.

    Labeling:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: The two Us in italics are not encoded by HTLV-1 and were added to facilitate fluorescent labeling and ensure free rotation of the dye. .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625.

    Purification:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: The purified protein was stored at −20 °C. .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625.

    Sequencing:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. In the final round of FACS, the upper 50% of mCherry-positive As-treated population was isolated for deep sequencing and identification of target genes.

    CRISPR:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: The genomic region containing the CRISPR guides was PCR-amplified and then subcloned into the lentiCRISPRv2 plasmid to generate a sublibrary. .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies).

    Plasmid Preparation:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625. .. A 15-nt 5′-extension (GGC CTT CGG GCC AAG) was added to these gRNA constructs to facilitate SHAPE analysis ( ).

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: .. The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies). .. Lentiviruses made from the sublibrary were then used to transduce into CHOP-mCherry reporter cells, which were subjected to a new round of As treatment and sorting.

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: .. Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. Similarly, to generate pcDNA 3.1-puro-mCherry, PCR amplified mCherry was cloned into EcoRI (New England Biolabs) linearized pcDNA 3.1-puro vector.

    In Vitro:

    Article Title: Human T-cell leukemia virus type 1 Gag domains have distinct RNA-binding specificities with implications for RNA packaging and dimerization
    Article Snippet: All other HTLV-1 gRNA-derived constructs were in vitro transcribed from digested plasmids using T7 RNA polymerase and previously established methods ( ). .. The following HTLV-1 gRNA constructs were derived from the proviral plasmid ACHneo ( ) (a gift from Patrick L. Green, Department of Veterinary Biosciences, The Ohio State University) and cloned into KpnI and PstI sites of pUC19 (New England Biolabs) in front of a T7 RNA polymerase promoter: HTLV-1 SL(447–512) derived from nt 447–512, R derived from nt 1–228, U5 derived from nt 229–450, 5′ gag derived from 450–625, and 5′ leader derived from nt 1–625.

    Marker:

    Article Title: Novel sub-cellular localizations and intra-molecular interactions may define new functions of Mixed Lineage Leukemia protein
    Article Snippet: Complementary DNA (cDNA) construct having full-length MLL was cloned into pBabe-FLAG vector as described [ ]. pcDNA 3.1-puro-GFP was made by inserting polymerase chain reaction (PCR) amplified GFP into KpnI (New England Biolabs) linearized pcDNA 3.1-puro vector. .. GFP-tagged pcDNA5/FRT and pcDNA5/FRT TO vector, having hygromycin as selectable marker, was generated by PCR sub-cloning of GFP into EcoRI and EcoRV (New England Biolabs) site of pcDNA5/FRT, Xho-1 site of pcDNA5/FRT TO vector respectively.

    FACS:

    Article Title: Genome-wide CRISPR screen identifies suppressors of endoplasmic reticulum stress-induced apoptosis
    Article Snippet: Paragraph title: FACS-Based Screen. ... The primers used were “forward: 5′-AACGGATCGGCACTGCGTGC and reverse: 5′- TGTGGGCGATGTGCGCTCTG.” For subcloning, the PCR products and lentiCRISPRv2 plasmid were digested using EcoR1 and KpnI (New England Biolabs) restriction enzymes and transformed into XL10-Gold Ultracompetent Cells (Agilent Technologies).

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  • 90
    New England Biolabs kpni hf
    Kpni Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kpni hf/product/New England Biolabs
    Average 90 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    kpni hf - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    90
    New England Biolabs kpn i
    RAD50 HS sites are not present in naïve CD4 T cells but are rapidly induced within 48 h during Th differentiation. ( A ) Naïve CD4 T cells from TCR-transgenic mice were subjected to DNase I HS analysis by using a probe hybridizing to the 12-kb 3′ <t>Kpn</t> I fragment of RAD50 . No HS sites were observed. As a control, the same membrane was rehybridized with a probe specific for the ≈21-kb Kpn I fragment containing the IL-13 gene and part of the IL-4 / IL-13 ). ( B ) DNA methylation/Southern blot analysis shows two regions of heavy cytosine-methylation in the 12-kb Kpn I fragment in naïve CD4 T cells (indicated by arrows). The regions coincide with the locations of HS sites RAD50-A and -B, respectively. ( C ) DNase I HS analysis on naïve CD4 T cells stimulated for 48 h with plate-bound anti-CD3/CD28 in the presence of IL-12 plus neutralizing antibodies to IL-4 ( Left ) or IL-4 cytokine plus neutralizing antibodies to IFN-γ ( Right ). Arrows indicate the appearance of HS sites under both stimulation conditions. ( D ) Same as in C except that naïve CD4 T cells were treated with cyclosporin A ( Right ) or ethanol control ( Left ) during the anti-CD3/CD28 and IL-4 cytokine stimulation.
    Kpn I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/kpn i/product/New England Biolabs
    Average 90 stars, based on 131 article reviews
    Price from $9.99 to $1999.99
    kpn i - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    Image Search Results


    RAD50 HS sites are not present in naïve CD4 T cells but are rapidly induced within 48 h during Th differentiation. ( A ) Naïve CD4 T cells from TCR-transgenic mice were subjected to DNase I HS analysis by using a probe hybridizing to the 12-kb 3′ Kpn I fragment of RAD50 . No HS sites were observed. As a control, the same membrane was rehybridized with a probe specific for the ≈21-kb Kpn I fragment containing the IL-13 gene and part of the IL-4 / IL-13 ). ( B ) DNA methylation/Southern blot analysis shows two regions of heavy cytosine-methylation in the 12-kb Kpn I fragment in naïve CD4 T cells (indicated by arrows). The regions coincide with the locations of HS sites RAD50-A and -B, respectively. ( C ) DNase I HS analysis on naïve CD4 T cells stimulated for 48 h with plate-bound anti-CD3/CD28 in the presence of IL-12 plus neutralizing antibodies to IL-4 ( Left ) or IL-4 cytokine plus neutralizing antibodies to IFN-γ ( Right ). Arrows indicate the appearance of HS sites under both stimulation conditions. ( D ) Same as in C except that naïve CD4 T cells were treated with cyclosporin A ( Right ) or ethanol control ( Left ) during the anti-CD3/CD28 and IL-4 cytokine stimulation.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Molecular analysis of a locus control region in the T helper 2 cytokine gene cluster: A target for STAT6 but not GATA3

    doi: 10.1073/pnas.0407031101

    Figure Lengend Snippet: RAD50 HS sites are not present in naïve CD4 T cells but are rapidly induced within 48 h during Th differentiation. ( A ) Naïve CD4 T cells from TCR-transgenic mice were subjected to DNase I HS analysis by using a probe hybridizing to the 12-kb 3′ Kpn I fragment of RAD50 . No HS sites were observed. As a control, the same membrane was rehybridized with a probe specific for the ≈21-kb Kpn I fragment containing the IL-13 gene and part of the IL-4 / IL-13 ). ( B ) DNA methylation/Southern blot analysis shows two regions of heavy cytosine-methylation in the 12-kb Kpn I fragment in naïve CD4 T cells (indicated by arrows). The regions coincide with the locations of HS sites RAD50-A and -B, respectively. ( C ) DNase I HS analysis on naïve CD4 T cells stimulated for 48 h with plate-bound anti-CD3/CD28 in the presence of IL-12 plus neutralizing antibodies to IL-4 ( Left ) or IL-4 cytokine plus neutralizing antibodies to IFN-γ ( Right ). Arrows indicate the appearance of HS sites under both stimulation conditions. ( D ) Same as in C except that naïve CD4 T cells were treated with cyclosporin A ( Right ) or ethanol control ( Left ) during the anti-CD3/CD28 and IL-4 cytokine stimulation.

    Article Snippet: Overlapping Bam HI and Kpn I (enzymes from New England Biolabs, Beverly, MA) genomic fragments spanning KIF3A to RAD50 were chosen for Southern blot analysis.

    Techniques: Transgenic Assay, Mouse Assay, DNA Methylation Assay, Southern Blot, Methylation

    Identification of previously unrecognized DNase I HS sites and regions of differential DNA methylation between Th1 and Th2 cells in the RAD50 gene. ( A ) Schematic diagram showing the IL-5 / RAD50 / IL-13 / IL-4 / KIF3A gene cluster. Shaded boxes represent genes, and arrows depict the direction of transcription. The gene regions indicated by bars B, C, and D are analyzed in B , C , and D , respectively. The gray bar above the locus indicates the extent of differential chromatin remodeling in Th1 and Th2 cells. ( B ) Southern analysis showing the positions of DNase I HS sites ( Left ) and regions of heavy DNA methylation ( Right ) within an ≈23-kb Bam HI region encompassing the KIF3A genes in D5 Th1 and D10 Th2 cells. ( C ) Same as in B except that the fragment analyzed is a 12-kb Kpn I fragment containing the very 3′ region of the RAD50 gene. Arrows indicate the common HS site, RAD50-A, and a Th2-specific HS site, RAD50-B. Both of these HS regions are heavily cytosine-methylated in Th1 but not in Th2 cells. ( D ) Same as in B except that the fragment analyzed is an ≈12-kb Bam HI fragment containing a central region of the RAD50 gene. The arrow indicates the common HS site RAD50-O.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Molecular analysis of a locus control region in the T helper 2 cytokine gene cluster: A target for STAT6 but not GATA3

    doi: 10.1073/pnas.0407031101

    Figure Lengend Snippet: Identification of previously unrecognized DNase I HS sites and regions of differential DNA methylation between Th1 and Th2 cells in the RAD50 gene. ( A ) Schematic diagram showing the IL-5 / RAD50 / IL-13 / IL-4 / KIF3A gene cluster. Shaded boxes represent genes, and arrows depict the direction of transcription. The gene regions indicated by bars B, C, and D are analyzed in B , C , and D , respectively. The gray bar above the locus indicates the extent of differential chromatin remodeling in Th1 and Th2 cells. ( B ) Southern analysis showing the positions of DNase I HS sites ( Left ) and regions of heavy DNA methylation ( Right ) within an ≈23-kb Bam HI region encompassing the KIF3A genes in D5 Th1 and D10 Th2 cells. ( C ) Same as in B except that the fragment analyzed is a 12-kb Kpn I fragment containing the very 3′ region of the RAD50 gene. Arrows indicate the common HS site, RAD50-A, and a Th2-specific HS site, RAD50-B. Both of these HS regions are heavily cytosine-methylated in Th1 but not in Th2 cells. ( D ) Same as in B except that the fragment analyzed is an ≈12-kb Bam HI fragment containing a central region of the RAD50 gene. The arrow indicates the common HS site RAD50-O.

    Article Snippet: Overlapping Bam HI and Kpn I (enzymes from New England Biolabs, Beverly, MA) genomic fragments spanning KIF3A to RAD50 were chosen for Southern blot analysis.

    Techniques: DNA Methylation Assay, Methylation