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bsiwi  (New England Biolabs)


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    Structured Review

    New England Biolabs bsiwi
    Bsiwi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 315 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/ecori-digested+lambda+vector/BsiWI/custom%40r0553%4033463551
    Average 96 stars, based on 315 article reviews
    bsiwi - by Bioz Stars, 2026-10
    96/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei
    Article Snippet: .. The neomycin resistance gene was PCR amplified from pC-PTP-NEO ( ) using primers UM184 and UM185 for subsequent Gibson assembly (NEB) with BsiWI and StuI digested pDEX-CuO ( ) to create pCuO-eGFP. ..

    Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei .
    Article Snippet: .. Cumate inducible expression vectors The neomycin resistance gene was PCR amplified from pC-PTP-NEO (24) using primers UM184 and UM185 for subsequent Gibson assembly (NEB) with BsiWI and StuI digested pDEX-CuO (21) to create pCuO-eGFP. ..

    Article Title: Epigenetic alterations facilitate transcriptional and translational programs in hypoxia.
    Article Snippet: The PDK1 ORF was amplified from pDONR223-PDK1 (Addgene, #23804) (primer pair PDK1_ORFstd_forw and PDK1_ORFstd_rev). .. A pGL4.13 plasmid (Promega, E6681), modified by introduction of a BsiWI restriction site downstream of the SV40 promoter, was digested with BsiWI (Pfl23II) and XbaI (Thermo Scientific, FD0854 and FD0684) to excise the 5′UTR and luciferase ORF, and PCR fragments containing the PDK1 5′UTR and ORF were inserted (NEBuilder HiFi DNA Assembly Cloning kit; NEB E5520S). .. To generate the CRISPR-resistant pGL4.13mod-134nt-PDK1-PAM, two PAM motifs within the PDK1 ORF were modified by site-directed mutagenesis (Platinum SuperFi PCR Master Mix; Thermo Scientific 12358010) with overlapping mutation-containing PCR primers (PDK1_ PAMmut_forw and PDK1_PAMmut_rev).

    Amplification:

    Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei
    Article Snippet: .. The neomycin resistance gene was PCR amplified from pC-PTP-NEO ( ) using primers UM184 and UM185 for subsequent Gibson assembly (NEB) with BsiWI and StuI digested pDEX-CuO ( ) to create pCuO-eGFP. ..

    Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei .
    Article Snippet: .. Cumate inducible expression vectors The neomycin resistance gene was PCR amplified from pC-PTP-NEO (24) using primers UM184 and UM185 for subsequent Gibson assembly (NEB) with BsiWI and StuI digested pDEX-CuO (21) to create pCuO-eGFP. ..

    Article Title: Rax1/2 promote memory of growth by early Ras1 activation to maintain polarity across generations
    Article Snippet: .. The amplified fragments were then assembled with either pFA6a-GFP-kanMX digested with BsiWI and SalI or pFA6a-mScarlet-natMX digested with PvuII and BamHI using the NEBuilder HiFi DNA Assembly Cloning Kit (#E5520S, New England Biolabs). ..

    Expressing:

    Article Title: A trypanosome trifecta: an independently tunable triple inducible system for genetic studies in Trypanosoma brucei .
    Article Snippet: .. Cumate inducible expression vectors The neomycin resistance gene was PCR amplified from pC-PTP-NEO (24) using primers UM184 and UM185 for subsequent Gibson assembly (NEB) with BsiWI and StuI digested pDEX-CuO (21) to create pCuO-eGFP. ..

    Plasmid Preparation:

    Article Title: In vivo -directed evolution identifies AAV-WM04 as a next-generation vector for potent and durable hearing restoration in DFNB9
    Article Snippet: .. The wild-type AAV2 capsid plasmid was linearized via restriction digestion with BsiWI (R0553V, NEB) and SnaBI (R0130V, NEB), flanking the insertion site. .. To construct the 9-mer library, a degenerate oligonucleotide encoding nine randomized amino acids (NNK codons) was synthesized (Suzhou GENEWIZ Biotechnology Co., Ltd.).

    Article Title: Leading and lagging strand abasic sites differentially affect vertebrate replisome progression but involve analogous bypass mechanisms
    Article Snippet: A plasmid designed to position the leading strand abasic site positioned 600 bp from the LacR array (pMC17) was commercially synthesized (Genscript) by inserting the oMC30/31 duplex in the reverse orientation at a position that places the damage site on the bottom strand 600 bp from the first lacO sequence of the array. .. To generate a plasmid containing a site-specific, but not strand-specific, AP site, pMC10 was digested with BsrGI and BsiWI (New England Biolabs) to remove the 50× LacR array. ..

    Article Title: Mapping the gene regulatory landscape of archaic hominin introgression in modern Papuans
    Article Snippet: .. The plasmid library was digested with BsiWI (NEB) and GFP (IDT) was cloned into the plasmid vector with HiFi assembly (NEBuilder HiFi DNA Assembly MasterMix). .. The final plasmid library was transformed into 10-beta electrocompetent E. coli (NEB) using a BioRad GenePulsar (2.0 kV, 200 Ω and 25 μF) and allowed to grow overnight in four flasks containing 500ml LB with ampicillin (final concentration = 0.1mg/ml).

    Article Title: Epigenetic alterations facilitate transcriptional and translational programs in hypoxia.
    Article Snippet: The PDK1 ORF was amplified from pDONR223-PDK1 (Addgene, #23804) (primer pair PDK1_ORFstd_forw and PDK1_ORFstd_rev). .. A pGL4.13 plasmid (Promega, E6681), modified by introduction of a BsiWI restriction site downstream of the SV40 promoter, was digested with BsiWI (Pfl23II) and XbaI (Thermo Scientific, FD0854 and FD0684) to excise the 5′UTR and luciferase ORF, and PCR fragments containing the PDK1 5′UTR and ORF were inserted (NEBuilder HiFi DNA Assembly Cloning kit; NEB E5520S). .. To generate the CRISPR-resistant pGL4.13mod-134nt-PDK1-PAM, two PAM motifs within the PDK1 ORF were modified by site-directed mutagenesis (Platinum SuperFi PCR Master Mix; Thermo Scientific 12358010) with overlapping mutation-containing PCR primers (PDK1_ PAMmut_forw and PDK1_PAMmut_rev).

    Clone Assay:

    Article Title: Mapping the gene regulatory landscape of archaic hominin introgression in modern Papuans
    Article Snippet: .. The plasmid library was digested with BsiWI (NEB) and GFP (IDT) was cloned into the plasmid vector with HiFi assembly (NEBuilder HiFi DNA Assembly MasterMix). .. The final plasmid library was transformed into 10-beta electrocompetent E. coli (NEB) using a BioRad GenePulsar (2.0 kV, 200 Ω and 25 μF) and allowed to grow overnight in four flasks containing 500ml LB with ampicillin (final concentration = 0.1mg/ml).

    Modification:

    Article Title: Epigenetic alterations facilitate transcriptional and translational programs in hypoxia.
    Article Snippet: The PDK1 ORF was amplified from pDONR223-PDK1 (Addgene, #23804) (primer pair PDK1_ORFstd_forw and PDK1_ORFstd_rev). .. A pGL4.13 plasmid (Promega, E6681), modified by introduction of a BsiWI restriction site downstream of the SV40 promoter, was digested with BsiWI (Pfl23II) and XbaI (Thermo Scientific, FD0854 and FD0684) to excise the 5′UTR and luciferase ORF, and PCR fragments containing the PDK1 5′UTR and ORF were inserted (NEBuilder HiFi DNA Assembly Cloning kit; NEB E5520S). .. To generate the CRISPR-resistant pGL4.13mod-134nt-PDK1-PAM, two PAM motifs within the PDK1 ORF were modified by site-directed mutagenesis (Platinum SuperFi PCR Master Mix; Thermo Scientific 12358010) with overlapping mutation-containing PCR primers (PDK1_ PAMmut_forw and PDK1_PAMmut_rev).

    Luciferase:

    Article Title: Epigenetic alterations facilitate transcriptional and translational programs in hypoxia.
    Article Snippet: The PDK1 ORF was amplified from pDONR223-PDK1 (Addgene, #23804) (primer pair PDK1_ORFstd_forw and PDK1_ORFstd_rev). .. A pGL4.13 plasmid (Promega, E6681), modified by introduction of a BsiWI restriction site downstream of the SV40 promoter, was digested with BsiWI (Pfl23II) and XbaI (Thermo Scientific, FD0854 and FD0684) to excise the 5′UTR and luciferase ORF, and PCR fragments containing the PDK1 5′UTR and ORF were inserted (NEBuilder HiFi DNA Assembly Cloning kit; NEB E5520S). .. To generate the CRISPR-resistant pGL4.13mod-134nt-PDK1-PAM, two PAM motifs within the PDK1 ORF were modified by site-directed mutagenesis (Platinum SuperFi PCR Master Mix; Thermo Scientific 12358010) with overlapping mutation-containing PCR primers (PDK1_ PAMmut_forw and PDK1_PAMmut_rev).

    Cloning:

    Article Title: Epigenetic alterations facilitate transcriptional and translational programs in hypoxia.
    Article Snippet: The PDK1 ORF was amplified from pDONR223-PDK1 (Addgene, #23804) (primer pair PDK1_ORFstd_forw and PDK1_ORFstd_rev). .. A pGL4.13 plasmid (Promega, E6681), modified by introduction of a BsiWI restriction site downstream of the SV40 promoter, was digested with BsiWI (Pfl23II) and XbaI (Thermo Scientific, FD0854 and FD0684) to excise the 5′UTR and luciferase ORF, and PCR fragments containing the PDK1 5′UTR and ORF were inserted (NEBuilder HiFi DNA Assembly Cloning kit; NEB E5520S). .. To generate the CRISPR-resistant pGL4.13mod-134nt-PDK1-PAM, two PAM motifs within the PDK1 ORF were modified by site-directed mutagenesis (Platinum SuperFi PCR Master Mix; Thermo Scientific 12358010) with overlapping mutation-containing PCR primers (PDK1_ PAMmut_forw and PDK1_PAMmut_rev).

    Article Title: Rax1/2 promote memory of growth by early Ras1 activation to maintain polarity across generations
    Article Snippet: .. The amplified fragments were then assembled with either pFA6a-GFP-kanMX digested with BsiWI and SalI or pFA6a-mScarlet-natMX digested with PvuII and BamHI using the NEBuilder HiFi DNA Assembly Cloning Kit (#E5520S, New England Biolabs). ..



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