nsii hf  (New England Biolabs)


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  • 95
    Name:
    NsiI HF
    Description:
    NsiI HF 5 000 units
    Catalog Number:
    R3127L
    Price:
    269
    Category:
    Restriction Enzymes
    Size:
    5 000 units
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    Structured Review

    New England Biolabs nsii hf
    NsiI HF
    NsiI HF 5 000 units
    https://www.bioz.com/result/nsii hf/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nsii hf - by Bioz Stars, 2021-05
    95/100 stars

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    Related Articles

    Concentration Assay:

    Article Title: A modular DNA scaffold to study protein-protein interactions at single-molecule resolution.
    Article Snippet: This reaction took place at a final concentration of ≈ 270 nM in each strand, for 1 h at room temperature in 1× CutSmart buffer (New England Biolabs). .. This assembly was subsequently combined with the leash precursor at ≈ 130 nM final concentration each in 1× CutSmart buffer, along with 10 units (U) of SbfI-HF, 10 U of MluI-HF, 10 U of NsiI-HF, 5 U of AscI, 1000 U of T4 DNA ligase (New England Biolabs), 1 mM ATP, and 1 mM DTT (Step a4 in Supp. .. After overnight incubation at 25 °C the mixture was heat inactivated at 65 °C for 20 min. Then, in a one pot reaction taking place at 37 °C for 8 - 16 h, the tips were nicked with 30 U of Nb.BvCI (New England Biolabs) and the extremities of the shanks were digested with 100 U of XbaI and 50 U of SacI (New England Biolabs) (Step a5 in Supp.

    Polymerase Chain Reaction:

    Article Title: Epstein-Barr virus BORF2 inhibits cellular APOBEC3B to preserve viral genome integrity
    Article Snippet: BORF2 with a C-terminal eGFP tag was generated by high-fidelity PCR of pcDNA4-BORF2-3xFlag using primers RSH13422 5’-NNN NAT GCA TCA TGG CAA CGA CCA GTC ATG TC-3’ and RSH13424 5’-NNN NAC GCG TCC TTG GCA AGA TTC ACA GGC TCG-3’. .. PCR products were digested with Nsi I-HF (NEB R3127) and Mlu I-HF (NEB R3198) and cloned into the previously described pQCXIP (Clontech) with a C-terminal eGFP tag . .. BORF2 with C-terminal 3x-Flag tag was cloned into an MLV-based pQCXIP lentivirus vector (Clontech) for complementation experiments by PCR of pcDNA4-BORF2-3xFlag using primers RSH13422 5’-NNN NAT GCA TCA TGG CAA CGA CCA GTC ATG TC-3’ and RSH13423 5’-NNN NTT AAT TAA TTA AAC GGG CCC CTT GTC GTC-3’.

    Clone Assay:

    Article Title: Epstein-Barr virus BORF2 inhibits cellular APOBEC3B to preserve viral genome integrity
    Article Snippet: BORF2 with a C-terminal eGFP tag was generated by high-fidelity PCR of pcDNA4-BORF2-3xFlag using primers RSH13422 5’-NNN NAT GCA TCA TGG CAA CGA CCA GTC ATG TC-3’ and RSH13424 5’-NNN NAC GCG TCC TTG GCA AGA TTC ACA GGC TCG-3’. .. PCR products were digested with Nsi I-HF (NEB R3127) and Mlu I-HF (NEB R3198) and cloned into the previously described pQCXIP (Clontech) with a C-terminal eGFP tag . .. BORF2 with C-terminal 3x-Flag tag was cloned into an MLV-based pQCXIP lentivirus vector (Clontech) for complementation experiments by PCR of pcDNA4-BORF2-3xFlag using primers RSH13422 5’-NNN NAT GCA TCA TGG CAA CGA CCA GTC ATG TC-3’ and RSH13423 5’-NNN NTT AAT TAA TTA AAC GGG CCC CTT GTC GTC-3’.

    Plasmid Preparation:

    Article Title: Development of a pan-neuronal genetic driver in Aedes aegypti mosquitoes
    Article Snippet: Two families were again positive at G2 and PCR followed by Sanger sequencing revealed proper genomic integration for one family. .. Syt1LeftArm-T2A-GAL4d-Hsp70-3XUAS-GCaMP6s-SV40-3XP3-dsRed-SV40-Syt1RightArm[1] Plasmid backbone from psL1180, linearized with restriction enzymes NsiI-HF (New England Biolabs #R3127S) and AvrII (New England Biolabs #R0174S). .. [2] GAL4d from pBPGAL4.1Uw (Addgene plasmid #26226)(Primers:ADdomain,forward,5’-caggcggccgccataTGCATGGATCCGCCAACTTC AACCAGAGTGG-3’, reverse, 5’-tatcgatagacgtca CTACTCCTTCTTTGGGTTCGG-3’; BD domain, forward,5’-ATGAAGCTGCTGAGTAGTATTGA AC-3’, reverse 5’-aagttggcggatccaGATACCGTC AGTTGCCGTTGAC-3’).

    Article Title: Development of a pan-neuronal genetic driver in Aedes aegypti mosquitoes
    Article Snippet: brpLeftArm-T2A-QF2w-Hsp70-3XP3-dsRed-SV40-brpRightArm[1] Plasmid backbone from psL1180, linearized with restriction enzymes NsiI-HF (New England Biolabs #R3127S) and AvrII (New England Biolabs #R0174S). .. brpLeftArm-T2A-QF2w-Hsp70-3XP3-dsRed-SV40-brpRightArm[1] Plasmid backbone from psL1180, linearized with restriction enzymes NsiI-HF (New England Biolabs #R3127S) and AvrII (New England Biolabs #R0174S). ..

    Article Title: Development of a pan-neuronal genetic driver in Aedes aegypti mosquitoes
    Article Snippet: All larvae from two families had strong green fluorescence in the central and peripheral nervous system, but died at the late larval stage. .. Syt1LeftArm-T2A-QF2-Hsp70-5XQUAS-GCaMP6s-SV40-3XP3-dsRed-SV40-Syt1RightArm [1] Plasmid backbone from psL1180, linearized with restriction enzyme NsiI-HF (New England Biolabs #R3127S). .. [2] QF2-Hsp70 from pattB-synaptobrevin-7-QFBDAD-hsp70 (Addgene plasmid #46115) (Primers: Forward, 5’-ATGCCACCCAAGCGCAA AACG-3’; Reverse, 5’-ccaagcttggatccaGGCCGC GGATCTAAACGAGTTT-3’).

    Article Title: A versatile dual-use RT-PCR control for use in assays for the detection of peste des petits ruminants virus.
    Article Snippet: The final PCR product was then gel purified (Invitrogen Clonewell II) and stored at -20°C prior to digestion. .. The insert was digested with NsiI-HF and AFlIII [New England Biolabs (NEB)] at 37°C for 4 hrs and then allowed to cool to room temperature (~20°C) at which point the digested template was gel purified (Invitrogen Clonewell II) and stored at -20°C prior to ligation. pBRT7Qβ plasmid DNA (Taniguchi et al., 1978) was digested with NsiI-HF and AFlIII (NEB) and dephosphorylated with FastAP (Thermo Scientific) at 4°C overnight followed by incubation for 3.0 hours at 37°C and heat inactivation at 80°C for 20 minutes. .. The PPRV control template was ligated into the 3’ end of the A1 gene of phage Qβ located on pBRT7Qβ in a 30.0 µL reaction containing 1.0 µL T4 ligase (NEB); 11.0 µL nuclease free water; 15.0 µL 2X ligation buffer (NEB); 1.0 µL digested pBRT7Qβ (29 ng) and 2.0 µL digested control insert (6.0 ng).

    Purification:

    Article Title: A versatile dual-use RT-PCR control for use in assays for the detection of peste des petits ruminants virus.
    Article Snippet: The final PCR product was then gel purified (Invitrogen Clonewell II) and stored at -20°C prior to digestion. .. The insert was digested with NsiI-HF and AFlIII [New England Biolabs (NEB)] at 37°C for 4 hrs and then allowed to cool to room temperature (~20°C) at which point the digested template was gel purified (Invitrogen Clonewell II) and stored at -20°C prior to ligation. pBRT7Qβ plasmid DNA (Taniguchi et al., 1978) was digested with NsiI-HF and AFlIII (NEB) and dephosphorylated with FastAP (Thermo Scientific) at 4°C overnight followed by incubation for 3.0 hours at 37°C and heat inactivation at 80°C for 20 minutes. .. The PPRV control template was ligated into the 3’ end of the A1 gene of phage Qβ located on pBRT7Qβ in a 30.0 µL reaction containing 1.0 µL T4 ligase (NEB); 11.0 µL nuclease free water; 15.0 µL 2X ligation buffer (NEB); 1.0 µL digested pBRT7Qβ (29 ng) and 2.0 µL digested control insert (6.0 ng).

    Ligation:

    Article Title: A versatile dual-use RT-PCR control for use in assays for the detection of peste des petits ruminants virus.
    Article Snippet: The final PCR product was then gel purified (Invitrogen Clonewell II) and stored at -20°C prior to digestion. .. The insert was digested with NsiI-HF and AFlIII [New England Biolabs (NEB)] at 37°C for 4 hrs and then allowed to cool to room temperature (~20°C) at which point the digested template was gel purified (Invitrogen Clonewell II) and stored at -20°C prior to ligation. pBRT7Qβ plasmid DNA (Taniguchi et al., 1978) was digested with NsiI-HF and AFlIII (NEB) and dephosphorylated with FastAP (Thermo Scientific) at 4°C overnight followed by incubation for 3.0 hours at 37°C and heat inactivation at 80°C for 20 minutes. .. The PPRV control template was ligated into the 3’ end of the A1 gene of phage Qβ located on pBRT7Qβ in a 30.0 µL reaction containing 1.0 µL T4 ligase (NEB); 11.0 µL nuclease free water; 15.0 µL 2X ligation buffer (NEB); 1.0 µL digested pBRT7Qβ (29 ng) and 2.0 µL digested control insert (6.0 ng).

    Incubation:

    Article Title: A versatile dual-use RT-PCR control for use in assays for the detection of peste des petits ruminants virus.
    Article Snippet: The final PCR product was then gel purified (Invitrogen Clonewell II) and stored at -20°C prior to digestion. .. The insert was digested with NsiI-HF and AFlIII [New England Biolabs (NEB)] at 37°C for 4 hrs and then allowed to cool to room temperature (~20°C) at which point the digested template was gel purified (Invitrogen Clonewell II) and stored at -20°C prior to ligation. pBRT7Qβ plasmid DNA (Taniguchi et al., 1978) was digested with NsiI-HF and AFlIII (NEB) and dephosphorylated with FastAP (Thermo Scientific) at 4°C overnight followed by incubation for 3.0 hours at 37°C and heat inactivation at 80°C for 20 minutes. .. The PPRV control template was ligated into the 3’ end of the A1 gene of phage Qβ located on pBRT7Qβ in a 30.0 µL reaction containing 1.0 µL T4 ligase (NEB); 11.0 µL nuclease free water; 15.0 µL 2X ligation buffer (NEB); 1.0 µL digested pBRT7Qβ (29 ng) and 2.0 µL digested control insert (6.0 ng).

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  • 95
    New England Biolabs nsii hf
    Nsii Hf, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nsii hf/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nsii hf - by Bioz Stars, 2021-05
    95/100 stars
      Buy from Supplier

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