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restriction enzymes pvu ii  (TaKaRa)


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    TaKaRa restriction enzymes pvu ii
    Restriction Enzymes Pvu Ii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 95/100, based on 488 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/restriction+enzymes+pvu+ii/Pvu+II/pmc12116657-128-9-28
    Average 95 stars, based on 488 article reviews
    restriction enzymes pvu ii - by Bioz Stars, 2026-10
    95/100 stars

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    other:

    Article Title: Gene structure and transcription of IRF-1 and IRF-7 in the mandarin fish Siniperca chuatsi.
    Article Snippet: The genes of IRF-1 and IRF-7 have been cloned from the mandarin fish (Siniperca chuatsi).. The IRF-1 gene has 4919 nucleotides (nt) and contains 10 exons and 9 introns, with an open reading frame (ORF) of 903 nt encoding 301 aa.. The IRF-7 gene has 6057 nt and also contains 10 exons and 9 introns, with an ORF of 1308 nt encoding 436 aa.

    Article Title: A “proto” type galectin expressed in striped bass ( Morone saxatilis ) tissues is released to epidermal mucus and binds to bacterial and mucus glycans
    Article Snippet: For this purpose, five genomic libraries were prepared using restriction enzymes Pvu II, Sca I, Eco RV, Stu I, and Dra I, ligated with the Genome Walker Adapter (Clontech) according to the manufacturer’s protocols.

    Polymerase Chain Reaction:

    Article Title: Cyclooxygenase-2 polymorphisms and the risk of gastric cancer in various degrees of relationship in the Chinese Han population
    Article Snippet: Thermo cycling conditions of PCR were: COX-2 -1195G>A: 95°C for 5 min, 35 cycles of 95°C for 30 sec, 54°C for 40 sec, 72 °C for 45 sec and finally 72°C for 10 min; COX-2 -765G>C: 95°C for 5 min, 35 cycles of 95°C s for 30 sec, 55.5°C for 40 sec, 72°C for 45 sec and finally 72°C for 10 min. .. The PCR products were digested with restriction enzymes Pvu II (Takara Biotechnology Co., Ltd.) at 37°C overnight for analysis of COX-2 -1195G>A and with Hha I (Takara Biotechnology Co., Ltd.) for -765G>C. ..



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    Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco <t>I,</t> <t>EcoR</t> I, Hind III , and <t>Pvu</t> II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .
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    Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco <t>I,</t> <t>EcoR</t> I, Hind III , and <t>Pvu</t> II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .
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    Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco <t>I,</t> <t>EcoR</t> I, Hind III , and <t>Pvu</t> II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .
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    Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco I, EcoR I, Hind III , and Pvu II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .

    Journal: BioMed Research International

    Article Title: Nonviral Gene Targeting at rDNA Locus of Human Mesenchymal Stem Cells

    doi: 10.1155/2013/135189

    Figure Lengend Snippet: Site-specific integration at the rDNA locus of MSCs. (a) Schematic of the construction of pHr2-NL. pHr2-NL contained two inverted expression cassettes, one consisting of an IRES element from the encephalomyocarditis virus, the coding region of the Neo gene, the SV40 polyA signal (SV40pA), and two loxP sites with the same orientation. LoxP sites were recognized by CRE enzyme to remove the Neo cassette after gene targeting. LHA, long homologous arm (U13369:937-6523); SHA, short homologous arm (U13369:6523–7643). The genomic locus indicates the 6.7 kb fragment (U13369:937-7643) required for homologous recombination at the internal transcribed spacer 1 (ITS1) of the rRNA gene. Single cutting sites for restricted enzymes of Nco I, EcoR I, Hind III , and Pvu II are located at the IRES-Neo frame and outside of the long homologous fragment. The fragment between the two Pvu II sites was 8285 bp in size, and it was detected using probe 1 (P1). The expected sizes of the restriction fragments produced by Nco I, EcoR I, and Hind III were 4001 bp, 7628 bp, and 15,316 bp, respectively. These were detected using probe 2 (P2). Primer t-up was located at the SV40 polyA. Primer t-re was located outside of the SHA at the hrDNA locus. (b) Drug-resistant cell in basal medium. (c) Drug-resistant colonies in the medium supplemented with VEGF+bFGF+Vc+ITS-X. (d) Identification of colonies with site-specific integration by PCR. The expected fragment, 1.3 kb in size, was amplified from the genomic DNA of colonies using site-specific integration. M, DL200 DNA marker; 1, negative colony; 2–5, positive colonies; 6, wild-type MSCs. (e–f) Southern blotting analysis of the representative recombinants. Genomic DNA digested with Pvu II , Nco I, EcoR I, and Hind III was analyzed. A specific band was consistently detected in colonies 1-1, 1-2, 2-1, and 2-2. An additional band beside the specific band was detected in colony 2-3. c, control (untransfected MSCs); N, Nco I; E, EcoR I; H, Hind III .

    Article Snippet: After overnight digestion with restriction enzymes Pvu II, Nco I, EcoR I, and Hind III (New England Biolabs, Ipswich, MA, USA), 3 μ g of genomic DNA per sample was electrophoresed on a 0.8% agarose gel overnight and then transferred to positively charged nylon membranes (Hybond-N+, Amersham, Piscataway, NJ, USA).

    Techniques: Expressing, Homologous Recombination, Produced, Amplification, Marker, Southern Blot