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  • 95
    New England Biolabs neb buffer1
    Neb Buffer1, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs nebuffer 1
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    New England Biolabs hpych4iv
    Hpych4iv, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 396 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs hpa ii
    Hpa Ii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 914 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs paci
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    99
    New England Biolabs kpn i
    Generation of LMP2A ITAM mutant transgenic mice. (A) LMP2A ITAM mutant transgene construct. The Eμ heavy-chain enhancer/promoter (wavy lines) was fused to the LMP2A ITAM mutant transgene containing both LMP2A cDNA sequences (striped lines) and EBV genomic sequences (stippled). The DNA, protein, and amino acid number (AA #) for the altered ITAM sequences are shown in the shaded inset at the bottom. A silent point mutation creates a <t>Kpn</t> I site in the LMP2A ITAM mutant transgene, which aids in distinguishing ITAM mutant mice from LMP2A transgenic mice by PCR using OL129 and OL131 (upper inset) followed by Kpn I digestion. The LMP2A cDNA probe shown spans sequences throughout the LMP2A transgene. Bam HI (B) sites are indicated. The Bam HI site in parentheses may be lost upon integration. (B) PCR/ Kpn I digest identification of LMP2A ITAM mutant transgenic mice. Tail genomic DNA sequences from LMP2A ITAM mutant, nonmutated LMP2A, and WT littermate control animals were amplified using OL129-OL131 and <t>RAG</t> positive control primers, followed by digestion with Kpn I (K) or no enzyme (NE). Identities of the amplified products and their subsequent digested fragments are shown to the left. φX DNA digested with Hae III was used as a size standard (right-most lane). (C) Southern hybridization analysis of LMP2A ITAM mutant lines. Tail DNAs from TgΔITAM1, TgΔITAM2, TgE, and WT animals were digested with Bam HI, run on an 0.8% agarose gel, and probed for LMP2A sequences by Southern hybridization. Size standards are indicated at the right.
    Kpn I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1341 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Generation of LMP2A ITAM mutant transgenic mice. (A) LMP2A ITAM mutant transgene construct. The Eμ heavy-chain enhancer/promoter (wavy lines) was fused to the LMP2A ITAM mutant transgene containing both LMP2A cDNA sequences (striped lines) and EBV genomic sequences (stippled). The DNA, protein, and amino acid number (AA #) for the altered ITAM sequences are shown in the shaded inset at the bottom. A silent point mutation creates a Kpn I site in the LMP2A ITAM mutant transgene, which aids in distinguishing ITAM mutant mice from LMP2A transgenic mice by PCR using OL129 and OL131 (upper inset) followed by Kpn I digestion. The LMP2A cDNA probe shown spans sequences throughout the LMP2A transgene. Bam HI (B) sites are indicated. The Bam HI site in parentheses may be lost upon integration. (B) PCR/ Kpn I digest identification of LMP2A ITAM mutant transgenic mice. Tail genomic DNA sequences from LMP2A ITAM mutant, nonmutated LMP2A, and WT littermate control animals were amplified using OL129-OL131 and RAG positive control primers, followed by digestion with Kpn I (K) or no enzyme (NE). Identities of the amplified products and their subsequent digested fragments are shown to the left. φX DNA digested with Hae III was used as a size standard (right-most lane). (C) Southern hybridization analysis of LMP2A ITAM mutant lines. Tail DNAs from TgΔITAM1, TgΔITAM2, TgE, and WT animals were digested with Bam HI, run on an 0.8% agarose gel, and probed for LMP2A sequences by Southern hybridization. Size standards are indicated at the right.

    Journal: Journal of Virology

    Article Title: The LMP2A ITAM Is Essential for Providing B Cells with Development and Survival Signals In Vivo

    doi:

    Figure Lengend Snippet: Generation of LMP2A ITAM mutant transgenic mice. (A) LMP2A ITAM mutant transgene construct. The Eμ heavy-chain enhancer/promoter (wavy lines) was fused to the LMP2A ITAM mutant transgene containing both LMP2A cDNA sequences (striped lines) and EBV genomic sequences (stippled). The DNA, protein, and amino acid number (AA #) for the altered ITAM sequences are shown in the shaded inset at the bottom. A silent point mutation creates a Kpn I site in the LMP2A ITAM mutant transgene, which aids in distinguishing ITAM mutant mice from LMP2A transgenic mice by PCR using OL129 and OL131 (upper inset) followed by Kpn I digestion. The LMP2A cDNA probe shown spans sequences throughout the LMP2A transgene. Bam HI (B) sites are indicated. The Bam HI site in parentheses may be lost upon integration. (B) PCR/ Kpn I digest identification of LMP2A ITAM mutant transgenic mice. Tail genomic DNA sequences from LMP2A ITAM mutant, nonmutated LMP2A, and WT littermate control animals were amplified using OL129-OL131 and RAG positive control primers, followed by digestion with Kpn I (K) or no enzyme (NE). Identities of the amplified products and their subsequent digested fragments are shown to the left. φX DNA digested with Hae III was used as a size standard (right-most lane). (C) Southern hybridization analysis of LMP2A ITAM mutant lines. Tail DNAs from TgΔITAM1, TgΔITAM2, TgE, and WT animals were digested with Bam HI, run on an 0.8% agarose gel, and probed for LMP2A sequences by Southern hybridization. Size standards are indicated at the right.

    Article Snippet: These primers were used, in combination with RAG internal control primers, in a PCR followed by overnight digestion with Kpn I (1× NEB buffer 1, 1× BSA, 25.0 μl of PCR mixture, 1.0 U of Kpn I [NEB]) at 37°C.

    Techniques: Mutagenesis, Transgenic Assay, Mouse Assay, Construct, Genomic Sequencing, Polymerase Chain Reaction, Amplification, Positive Control, Hybridization, Agarose Gel Electrophoresis