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  • 99
    Millipore bgl ii bam hi sites
    Identification of yeast CUP1 transgene by southern blot and western blot analysis. (A) Southern blot analysis of the transgenes. Purified genomic <t>DNA</t> from each transgenic founder was digested with <t>Bgl</t> II and Ssp I, and analyzed by southern blotting using probes specific for CUP1 . Transgenic founders numbered 5, 6, 15, 20, 22, and 26; N: genomic DNA of control mice as a negative control. (B) Western blot was performed to confirm expression of CUP1 in the parotid and submandibular glands and in the saliva of transgenic lines. The expected protein size is 9 kDa. The lower band is β-actin (42 kDa, used as an internal control). The results demonstrated high expression of CUP1 in the parotid and submandibular glands after normalization against β-actin, as well as that in the saliva of transgenic mice. PG1 and PG2: the parotid glands of transgenic mice; SG1 and SG2: the submandibular glands of transgenic mice; Sa: the saliva of transgenic mice; N1, N2, and N3: the parotid gland, the submandibular gland and the saliva of control mice as negative controls, respectively.
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    99
    TaKaRa bgl ii cassette
    Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, <t>Bcl</t> I, <t>Bgl</t> l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.
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    93
    Bio-Rad bgl ii
    The bat and heb phenotypes result from mutation of Frem1 . ( A ) Sequencing of all 36 Frem1 coding exons identified an ENU induced T-C mutation in the splice donor site of exon 25 ( Left ). RT-PCR analysis shows that this mutation leads to skipping of exon 25 in bat RNA compared with C57BL/6J mice on which the mutation was induced. ( B ) Southern blot analysis of heb <t>DNA</t> with a probe spanning exon 17 detected a genomic rearrangement not present in the parental AKR/J strain. Inverse PCR identified the insertion of a LINE1 element 41 bp from the end of the exon. The <t>Bgl</t> II sites and primers used for inverse PCR (Pr, Pf, Dr, and Df) are indicated. ( C ) Position of the heb and bat mutations relative to the protein domains of Frem1 (CSPG, light gray; Calx, black; LectinC, dark gray).
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    Image Search Results


    Identification of yeast CUP1 transgene by southern blot and western blot analysis. (A) Southern blot analysis of the transgenes. Purified genomic DNA from each transgenic founder was digested with Bgl II and Ssp I, and analyzed by southern blotting using probes specific for CUP1 . Transgenic founders numbered 5, 6, 15, 20, 22, and 26; N: genomic DNA of control mice as a negative control. (B) Western blot was performed to confirm expression of CUP1 in the parotid and submandibular glands and in the saliva of transgenic lines. The expected protein size is 9 kDa. The lower band is β-actin (42 kDa, used as an internal control). The results demonstrated high expression of CUP1 in the parotid and submandibular glands after normalization against β-actin, as well as that in the saliva of transgenic mice. PG1 and PG2: the parotid glands of transgenic mice; SG1 and SG2: the submandibular glands of transgenic mice; Sa: the saliva of transgenic mice; N1, N2, and N3: the parotid gland, the submandibular gland and the saliva of control mice as negative controls, respectively.

    Journal: PLoS ONE

    Article Title: Transgenic Mice Expressing Yeast CUP1 Exhibit Increased Copper Utilization from Feeds

    doi: 10.1371/journal.pone.0107810

    Figure Lengend Snippet: Identification of yeast CUP1 transgene by southern blot and western blot analysis. (A) Southern blot analysis of the transgenes. Purified genomic DNA from each transgenic founder was digested with Bgl II and Ssp I, and analyzed by southern blotting using probes specific for CUP1 . Transgenic founders numbered 5, 6, 15, 20, 22, and 26; N: genomic DNA of control mice as a negative control. (B) Western blot was performed to confirm expression of CUP1 in the parotid and submandibular glands and in the saliva of transgenic lines. The expected protein size is 9 kDa. The lower band is β-actin (42 kDa, used as an internal control). The results demonstrated high expression of CUP1 in the parotid and submandibular glands after normalization against β-actin, as well as that in the saliva of transgenic mice. PG1 and PG2: the parotid glands of transgenic mice; SG1 and SG2: the submandibular glands of transgenic mice; Sa: the saliva of transgenic mice; N1, N2, and N3: the parotid gland, the submandibular gland and the saliva of control mice as negative controls, respectively.

    Article Snippet: Twenty micrograms of DNA was restriction digested with Bgl II and Ssp I, fractionated in a 0.8% agarose gel electrophoresis, and transferred to a nylon membrane (Millipore, UK).

    Techniques: Southern Blot, Western Blot, Purification, Transgenic Assay, Mouse Assay, Negative Control, Expressing

    Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Journal: Emerging Microbes & Infections

    Article Title: Comparative genomic analysis of two emergent human adenovirus type 14 respiratory pathogen isolates in China reveals similar yet divergent genomes

    doi: 10.1038/emi.2017.78

    Figure Lengend Snippet: Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Article Snippet: Genome type identification In silico restriction maps of available genome sequences from all HAdV-14 strains CHN/GZ01/2010, 303600, CHN/BJ430, CHN2012, and prototype de Wit were generated using the Vector NTI 10.3.0 software (Invitrogen Corp.; San Diego, CA, USA) as described in earlier studies., , These included profiles from restriction enzymes Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal l, Sma I, Xba I, and Xho I (TaKaRa Corp.; China), all of which were chosen in order to be consistent with the original nomenclature system devised by Li and colleagues.

    Techniques: In Silico, Generated, Plasmid Preparation, Software

    In vitro MMR efficiency of BS cell lines. ( A ) Western blot showing the absence of BLM protein in the BS cell lines. Aliquots of 25 µg of the indicated nuclear extracts were probed with anti-BLM antibody (IHIC33) and anti-XPB antibody [TFIIH p89 (S-19); Santa Cruz Biotechnology] as control. ( B ) MMR efficiency of the BLM-negative human fibroblast GM08505 and of the human lymphoblasts GM09960 and GM03403. The MMR-proficient MRC5 SV40 fibroblasts, TK6 lymphoblasts, PSNF5 cells (GM08505 stably transfected with BLM cDNA) and the MMR-deficient hMLH1 –/– colon cancer cell line HCT116 were used as controls. The MMR efficiencies of the two BS lymphoblasts complemented with purified recombinant BLM protein are also shown. The repair efficiency is expressed as fmol phagemid DNA cleaved by Bgl II in 30 min.

    Journal: Nucleic Acids Research

    Article Title: Direct association of Bloom's syndrome gene product with the human mismatch repair protein MLH1

    doi:

    Figure Lengend Snippet: In vitro MMR efficiency of BS cell lines. ( A ) Western blot showing the absence of BLM protein in the BS cell lines. Aliquots of 25 µg of the indicated nuclear extracts were probed with anti-BLM antibody (IHIC33) and anti-XPB antibody [TFIIH p89 (S-19); Santa Cruz Biotechnology] as control. ( B ) MMR efficiency of the BLM-negative human fibroblast GM08505 and of the human lymphoblasts GM09960 and GM03403. The MMR-proficient MRC5 SV40 fibroblasts, TK6 lymphoblasts, PSNF5 cells (GM08505 stably transfected with BLM cDNA) and the MMR-deficient hMLH1 –/– colon cancer cell line HCT116 were used as controls. The MMR efficiencies of the two BS lymphoblasts complemented with purified recombinant BLM protein are also shown. The repair efficiency is expressed as fmol phagemid DNA cleaved by Bgl II in 30 min.

    Article Snippet: The yeast strain L40 [ MATa trp1 leu2 his3 LYS2 :: lexA-HIS3 URA3 :: lexA-lacZ ] was sequentially transformed with the bait pBTM116-BLM (amino acids 770–1417) and a random primed human peripheral blood cDNA library cloned into the Bgl II sites of pACT (Clonetech) using the lithium acetate method.

    Techniques: In Vitro, Western Blot, Stable Transfection, Transfection, Purification, Recombinant

    The bat and heb phenotypes result from mutation of Frem1 . ( A ) Sequencing of all 36 Frem1 coding exons identified an ENU induced T-C mutation in the splice donor site of exon 25 ( Left ). RT-PCR analysis shows that this mutation leads to skipping of exon 25 in bat RNA compared with C57BL/6J mice on which the mutation was induced. ( B ) Southern blot analysis of heb DNA with a probe spanning exon 17 detected a genomic rearrangement not present in the parental AKR/J strain. Inverse PCR identified the insertion of a LINE1 element 41 bp from the end of the exon. The Bgl II sites and primers used for inverse PCR (Pr, Pf, Dr, and Df) are indicated. ( C ) Position of the heb and bat mutations relative to the protein domains of Frem1 (CSPG, light gray; Calx, black; LectinC, dark gray).

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

    Article Title: The extracellular matrix gene Frem1 is essential for the normal adhesion of the embryonic epidermis

    doi: 10.1073/pnas.0402760101

    Figure Lengend Snippet: The bat and heb phenotypes result from mutation of Frem1 . ( A ) Sequencing of all 36 Frem1 coding exons identified an ENU induced T-C mutation in the splice donor site of exon 25 ( Left ). RT-PCR analysis shows that this mutation leads to skipping of exon 25 in bat RNA compared with C57BL/6J mice on which the mutation was induced. ( B ) Southern blot analysis of heb DNA with a probe spanning exon 17 detected a genomic rearrangement not present in the parental AKR/J strain. Inverse PCR identified the insertion of a LINE1 element 41 bp from the end of the exon. The Bgl II sites and primers used for inverse PCR (Pr, Pf, Dr, and Df) are indicated. ( C ) Position of the heb and bat mutations relative to the protein domains of Frem1 (CSPG, light gray; Calx, black; LectinC, dark gray).

    Article Snippet: To identify the heb rearrangement, 5 μg of DNA from heb and AKR/J mice (The Jackson Laboratory) was digested with Bgl II and Southern blotted onto Zeta-Probe GT genomic membranes (Bio-Rad).

    Techniques: Mutagenesis, Sequencing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Southern Blot, Inverse PCR