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restriction enzyme cloning with xbai  (New England Biolabs)


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    Structured Review

    New England Biolabs restriction enzyme cloning with xbai
    Restriction Enzyme Cloning With Xbai, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 6783 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/restriction+enzyme+cloning+with+xbai/XbaI/pm41714755-141-33-38
    Average 99 stars, based on 6783 article reviews
    restriction enzyme cloning with xbai - by Bioz Stars, 2026-09
    99/100 stars

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    Related Articles

    Expressing:

    Article Title: Functional effects of EpCAM N-glycosylation in MDA-MB-468 breast cancer cells.
    Article Snippet: Generation of stable cell lines HEK293T cells were transfected with pLentiCRISPRv229 with EpCAM guide RNA30 (Genscript, Piscataway, NJ, USA), lentiviral packaging plasmid psPAX2 (courtesy of Didier Trono, Addgene #12260, Watertown, MA, USA), and envelope plasmid PMD2.G (courtesy of Didier Trono, Addgene #12259) using Lipofectamine 3000 (Thermo Fisher Scientific #L3000-008) according to the manufacturer’s instructions. .. To generate the EpCAM overexpressing (OV) and mutEpCAM expressing plasmids, custom genes were produced by Thermo Fisher Scientific GeneArt and inserted into pLentiCMV-blast31 (courtesy of Eric Campeau & Paul Kaufman, Addgene #17486) using restriction enzyme cloning with XbaI (New England Biolabs #R0145S) and SalI-HF (New England Biolabs #R3138S) using NEB rCutSmart Buffer (New England Biolabs #B6004S). .. Fragments were isolated by agarose (VWR #490001-580) gel electrophoresis as described above and purified using QIAquick PCR & Gel Cleanup kit (Qiagen #28506).

    Produced:

    Article Title: Functional effects of EpCAM N-glycosylation in MDA-MB-468 breast cancer cells.
    Article Snippet: Generation of stable cell lines HEK293T cells were transfected with pLentiCRISPRv229 with EpCAM guide RNA30 (Genscript, Piscataway, NJ, USA), lentiviral packaging plasmid psPAX2 (courtesy of Didier Trono, Addgene #12260, Watertown, MA, USA), and envelope plasmid PMD2.G (courtesy of Didier Trono, Addgene #12259) using Lipofectamine 3000 (Thermo Fisher Scientific #L3000-008) according to the manufacturer’s instructions. .. To generate the EpCAM overexpressing (OV) and mutEpCAM expressing plasmids, custom genes were produced by Thermo Fisher Scientific GeneArt and inserted into pLentiCMV-blast31 (courtesy of Eric Campeau & Paul Kaufman, Addgene #17486) using restriction enzyme cloning with XbaI (New England Biolabs #R0145S) and SalI-HF (New England Biolabs #R3138S) using NEB rCutSmart Buffer (New England Biolabs #B6004S). .. Fragments were isolated by agarose (VWR #490001-580) gel electrophoresis as described above and purified using QIAquick PCR & Gel Cleanup kit (Qiagen #28506).

    Cloning:

    Article Title: Functional effects of EpCAM N-glycosylation in MDA-MB-468 breast cancer cells.
    Article Snippet: Generation of stable cell lines HEK293T cells were transfected with pLentiCRISPRv229 with EpCAM guide RNA30 (Genscript, Piscataway, NJ, USA), lentiviral packaging plasmid psPAX2 (courtesy of Didier Trono, Addgene #12260, Watertown, MA, USA), and envelope plasmid PMD2.G (courtesy of Didier Trono, Addgene #12259) using Lipofectamine 3000 (Thermo Fisher Scientific #L3000-008) according to the manufacturer’s instructions. .. To generate the EpCAM overexpressing (OV) and mutEpCAM expressing plasmids, custom genes were produced by Thermo Fisher Scientific GeneArt and inserted into pLentiCMV-blast31 (courtesy of Eric Campeau & Paul Kaufman, Addgene #17486) using restriction enzyme cloning with XbaI (New England Biolabs #R0145S) and SalI-HF (New England Biolabs #R3138S) using NEB rCutSmart Buffer (New England Biolabs #B6004S). .. Fragments were isolated by agarose (VWR #490001-580) gel electrophoresis as described above and purified using QIAquick PCR & Gel Cleanup kit (Qiagen #28506).

    other:

    Article Title: Functional effects of EpCAM N-glycosylation in MDA-MB-468 breast cancer cells
    Article Snippet: After puromycin selection and a round of cell sorting with an Aria IIu Cell Sorter the Johns Hopkins Ross Flow Cytometry Core Facility using an anti-EpCAM antibody conjugated to Alexa Fluor 647 (1:200 dilution, Santa Cruz Biotechnology #sc25308-AF647, Dallas, TX, USA), the EpCAM knockout (KO) remained unstable, showing EpCAM expression to slowly increase again over time, indicating a mixed EpCAM KO and WT cell population.



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