nhei  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier
Bioz Manufacturer Symbol New England Biolabs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Name:
    NheI HF
    Description:
    NheI HF 5 000 units
    Catalog Number:
    r3131l
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    5 000 units
    Buy from Supplier


    Structured Review

    New England Biolabs nhei
    NheI HF
    NheI HF 5 000 units
    https://www.bioz.com/result/nhei/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nhei - by Bioz Stars, 2021-03
    95/100 stars

    Images

    Related Articles

    Amplification:

    Article Title: Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems
    Article Snippet: To clone RiboJ::sfGFP into the plasmid, RiboJ::sfGFP was first amplified with NEBNext® Q5® Hot Start HiFi PCR Master Mix (#M0543S) for 25 cycles using primers GU 99 and GU 100 at 10 µM. .. This amplicon was then digested with BsaI-HF (NEB # R3535) and NcoI-HF (NEB #R3193S) for 1.5 h at 37 °C. pLib was digested with BsaI-HF (NEB # R3535) and NheI (NEB# R3131S). pLib vector was then ligated with the GFP insert using T7 DNA Ligase (NEB #M0318S), incubating at room temperature for 1 h. This plasmid was next transformed into DH5α electrocompetent cells and plated for 24 h of growth at 30 °C as well, yielding pLib_sfGFP plasmid after maxi-prep. .. Library integration The pLib_sfGFP plasmid was first digested with SalI-HF (NEB #R3138S) and NheI (NEB# R3131S) to remove the background.

    Article Title: Capturing and Recreating Diverse Antibody Repertoires as Multivalent Recombinant Polyclonal Antibody Drugs
    Article Snippet: Plasmid purification was performed using ZymoPURE II Plasmid Maxiprep Kits (Zymo Research, Irvine, CA, USA). .. To create the full-length antibody library, we performed a second Gibson assembly by linearizing the product of GA1 with BamHI-HF (rHIG) or NheI-HF (rCIG, rPIG, rhATG, and rZIG) (NEB, Ipswich, MA, USA) and using it as a vector to insert a synthetic amplicon containing a portion of the light chain Ig constant region, a poly(A) signal for light chain Ig, a promoter for the IgG gene and a secretory leader sequence for the IgG gene. .. The full-length library was then transformed into E. coli and spread on LB-ampicillin plates.

    Plasmid Preparation:

    Article Title: Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems
    Article Snippet: To clone RiboJ::sfGFP into the plasmid, RiboJ::sfGFP was first amplified with NEBNext® Q5® Hot Start HiFi PCR Master Mix (#M0543S) for 25 cycles using primers GU 99 and GU 100 at 10 µM. .. This amplicon was then digested with BsaI-HF (NEB # R3535) and NcoI-HF (NEB #R3193S) for 1.5 h at 37 °C. pLib was digested with BsaI-HF (NEB # R3535) and NheI (NEB# R3131S). pLib vector was then ligated with the GFP insert using T7 DNA Ligase (NEB #M0318S), incubating at room temperature for 1 h. This plasmid was next transformed into DH5α electrocompetent cells and plated for 24 h of growth at 30 °C as well, yielding pLib_sfGFP plasmid after maxi-prep. .. Library integration The pLib_sfGFP plasmid was first digested with SalI-HF (NEB #R3138S) and NheI (NEB# R3131S) to remove the background.

    Article Title: Capturing and Recreating Diverse Antibody Repertoires as Multivalent Recombinant Polyclonal Antibody Drugs
    Article Snippet: Plasmid purification was performed using ZymoPURE II Plasmid Maxiprep Kits (Zymo Research, Irvine, CA, USA). .. To create the full-length antibody library, we performed a second Gibson assembly by linearizing the product of GA1 with BamHI-HF (rHIG) or NheI-HF (rCIG, rPIG, rhATG, and rZIG) (NEB, Ipswich, MA, USA) and using it as a vector to insert a synthetic amplicon containing a portion of the light chain Ig constant region, a poly(A) signal for light chain Ig, a promoter for the IgG gene and a secretory leader sequence for the IgG gene. .. The full-length library was then transformed into E. coli and spread on LB-ampicillin plates.

    Article Title: Nonspecificity in a nonimmune human scFv repertoire
    Article Snippet: The fragment before and containing the H2 was amplified with the pCTFwd Recomb and appropriate PreH2 primers, and the fragment after and also containing the H2 was amplified using the pcTRev Recomb and proper PostH2 primers (Primer sequences in Supplementary Table 1). .. The pCTCON2 vector was prepared for homologous recombination by digestion with SalI-HF followed by digestion with NheI-HF and BamI-HF (New England Biolabs). .. The two scFv fragments along with the digested vector were contransformed into chemically competent yeast using the Frozen-EZ Yeast Transformation II Kit (Zymo Research) and resultant clones were sequence verified before experiments.

    Article Title: Tango1 spatially organizes ER exit sites to control ER export
    Article Snippet: This produced pTG-NheI-G-XhoI-SpeI-W, which contains two copies of GFP. .. The original GFP sequence in this plasmid was then excised by double digestion with XbaI (cat#R0145S; New England Biolabs, Inc.), for which a restriction site was already present between the UAS promoter and GFP in the original pTGW, and NheI (catR3131L; New England Biolabs, Inc.). .. The excised GFP sequence was replaced through restriction and ligation with T4 DNA ligase (cat#M0202L; New England Biolabs, Inc.) by the signal peptide of Tango1 (26 first amino acids, predicted by SignalP 4.1), which we PCR-amplified from pDONR221-Tango1 using primers that added the appropriate restriction sites (XbaISP-F: 5′-GGCTCTAGAATGCGGCTGACCAACGAGA-3′ and NheISP-R: 5′-CTAGCTAGCAGCCCACGTCAAAGTTGGAA-3′).

    Article Title: Directed evolution of broadly crossreactive chemokine-blocking antibodies efficacious in arthritis
    Article Snippet: To ensure a mutagenesis rate of approximately 1–2 amino-acid mutated residues distributed randomly throughout the entire gene, 1 ng of DNA template encoding the CK1, CK2, and CK4 binders were PCR amplified for 15 cycles using Taq DNA polymerase (New England BioLabs), analog nucleotides (2 μM 8-oxo-dGTP and 2 μM dPTP) and flanking oligonucleotide primers (forward: 5′-GGAGGCGGTAGCGGAGGCGGAGGGTCGGCTAGC-3′; reverse: 5′-GTCCTCTTCAGAAATAAGCTTTTGTTCGGAT-3′; Integrated DNA Technologies). .. The mutagenized PCR products were further purified, re-amplified for additional 30 cycles in the absence of analog nucleotides, and combined with Sal I-HF, Nhe I-HF, and Bam HI-HI (New England BioLabs) digested pCT-CON vector. .. Pre-mixed DNA linearized vector and PCR insert (1 μg μL−1 ) was electroporated into freshly prepare EBY100 competent cells, where the full constructs are reassembled via homologous recombination .

    Transformation Assay:

    Article Title: Multiplexed characterization of rationally designed promoter architectures deconstructs combinatorial logic for IPTG-inducible systems
    Article Snippet: To clone RiboJ::sfGFP into the plasmid, RiboJ::sfGFP was first amplified with NEBNext® Q5® Hot Start HiFi PCR Master Mix (#M0543S) for 25 cycles using primers GU 99 and GU 100 at 10 µM. .. This amplicon was then digested with BsaI-HF (NEB # R3535) and NcoI-HF (NEB #R3193S) for 1.5 h at 37 °C. pLib was digested with BsaI-HF (NEB # R3535) and NheI (NEB# R3131S). pLib vector was then ligated with the GFP insert using T7 DNA Ligase (NEB #M0318S), incubating at room temperature for 1 h. This plasmid was next transformed into DH5α electrocompetent cells and plated for 24 h of growth at 30 °C as well, yielding pLib_sfGFP plasmid after maxi-prep. .. Library integration The pLib_sfGFP plasmid was first digested with SalI-HF (NEB #R3138S) and NheI (NEB# R3131S) to remove the background.

    Purification:

    Article Title: Transcriptional and posttranscriptional regulation of Shigella shuT in response to host‐associated iron availability and temperature. Transcriptional and posttranscriptional regulation of Shigella shuT in response to host‐associated iron availability and temperature
    Article Snippet: 2.12.2 In vitro transcription E. coli strain DH5α carrying the run‐off plasmid pT7‐T was cultured to the stationary phase of growth in 3 ml LB broth with 30 (μg ml−1 ) chloramphenicol at 37°C. .. Following extraction, purified pT7‐T was linearized by digestion with restriction enzyme NheI‐HF (New England Biolabs Inc.) at 37°C for 2 hr. .. After purification by gel extraction, the linear DNA was used as template in an in vitro transcription reaction carried out according to the protocol provided by AmpliScribe™ T7‐Flash™ Transcription kit (epicenter, an Illumina company).

    Article Title: Directed evolution of broadly crossreactive chemokine-blocking antibodies efficacious in arthritis
    Article Snippet: To ensure a mutagenesis rate of approximately 1–2 amino-acid mutated residues distributed randomly throughout the entire gene, 1 ng of DNA template encoding the CK1, CK2, and CK4 binders were PCR amplified for 15 cycles using Taq DNA polymerase (New England BioLabs), analog nucleotides (2 μM 8-oxo-dGTP and 2 μM dPTP) and flanking oligonucleotide primers (forward: 5′-GGAGGCGGTAGCGGAGGCGGAGGGTCGGCTAGC-3′; reverse: 5′-GTCCTCTTCAGAAATAAGCTTTTGTTCGGAT-3′; Integrated DNA Technologies). .. The mutagenized PCR products were further purified, re-amplified for additional 30 cycles in the absence of analog nucleotides, and combined with Sal I-HF, Nhe I-HF, and Bam HI-HI (New England BioLabs) digested pCT-CON vector. .. Pre-mixed DNA linearized vector and PCR insert (1 μg μL−1 ) was electroporated into freshly prepare EBY100 competent cells, where the full constructs are reassembled via homologous recombination .

    Sequencing:

    Article Title: Capturing and Recreating Diverse Antibody Repertoires as Multivalent Recombinant Polyclonal Antibody Drugs
    Article Snippet: Plasmid purification was performed using ZymoPURE II Plasmid Maxiprep Kits (Zymo Research, Irvine, CA, USA). .. To create the full-length antibody library, we performed a second Gibson assembly by linearizing the product of GA1 with BamHI-HF (rHIG) or NheI-HF (rCIG, rPIG, rhATG, and rZIG) (NEB, Ipswich, MA, USA) and using it as a vector to insert a synthetic amplicon containing a portion of the light chain Ig constant region, a poly(A) signal for light chain Ig, a promoter for the IgG gene and a secretory leader sequence for the IgG gene. .. The full-length library was then transformed into E. coli and spread on LB-ampicillin plates.

    Article Title: Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization
    Article Snippet: Homology arms were inserted into pUC19 using an In-Fusion HD Cloning Kit. .. After confirmation of the sequence, homology arms and rTyr -repair cassette were excised using restriction endonucleases, Nhe I-HF and Mlu I-HF (New England Biolabs, Japan, Tokyo, Japan), and then ligated with pAAV_MCS2. ..

    Article Title: Tango1 spatially organizes ER exit sites to control ER export
    Article Snippet: This produced pTG-NheI-G-XhoI-SpeI-W, which contains two copies of GFP. .. The original GFP sequence in this plasmid was then excised by double digestion with XbaI (cat#R0145S; New England Biolabs, Inc.), for which a restriction site was already present between the UAS promoter and GFP in the original pTGW, and NheI (catR3131L; New England Biolabs, Inc.). .. The excised GFP sequence was replaced through restriction and ligation with T4 DNA ligase (cat#M0202L; New England Biolabs, Inc.) by the signal peptide of Tango1 (26 first amino acids, predicted by SignalP 4.1), which we PCR-amplified from pDONR221-Tango1 using primers that added the appropriate restriction sites (XbaISP-F: 5′-GGCTCTAGAATGCGGCTGACCAACGAGA-3′ and NheISP-R: 5′-CTAGCTAGCAGCCCACGTCAAAGTTGGAA-3′).

    Homologous Recombination:

    Article Title: Nonspecificity in a nonimmune human scFv repertoire
    Article Snippet: The fragment before and containing the H2 was amplified with the pCTFwd Recomb and appropriate PreH2 primers, and the fragment after and also containing the H2 was amplified using the pcTRev Recomb and proper PostH2 primers (Primer sequences in Supplementary Table 1). .. The pCTCON2 vector was prepared for homologous recombination by digestion with SalI-HF followed by digestion with NheI-HF and BamI-HF (New England Biolabs). .. The two scFv fragments along with the digested vector were contransformed into chemically competent yeast using the Frozen-EZ Yeast Transformation II Kit (Zymo Research) and resultant clones were sequence verified before experiments.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Tango1 spatially organizes ER exit sites to control ER export
    Article Snippet: This produced pTG-NheI-G-XhoI-SpeI-W, which contains two copies of GFP. .. The original GFP sequence in this plasmid was then excised by double digestion with XbaI (cat#R0145S; New England Biolabs, Inc.), for which a restriction site was already present between the UAS promoter and GFP in the original pTGW, and NheI (catR3131L; New England Biolabs, Inc.). .. The excised GFP sequence was replaced through restriction and ligation with T4 DNA ligase (cat#M0202L; New England Biolabs, Inc.) by the signal peptide of Tango1 (26 first amino acids, predicted by SignalP 4.1), which we PCR-amplified from pDONR221-Tango1 using primers that added the appropriate restriction sites (XbaISP-F: 5′-GGCTCTAGAATGCGGCTGACCAACGAGA-3′ and NheISP-R: 5′-CTAGCTAGCAGCCCACGTCAAAGTTGGAA-3′).

    Polymerase Chain Reaction:

    Article Title: Directed evolution of broadly crossreactive chemokine-blocking antibodies efficacious in arthritis
    Article Snippet: To ensure a mutagenesis rate of approximately 1–2 amino-acid mutated residues distributed randomly throughout the entire gene, 1 ng of DNA template encoding the CK1, CK2, and CK4 binders were PCR amplified for 15 cycles using Taq DNA polymerase (New England BioLabs), analog nucleotides (2 μM 8-oxo-dGTP and 2 μM dPTP) and flanking oligonucleotide primers (forward: 5′-GGAGGCGGTAGCGGAGGCGGAGGGTCGGCTAGC-3′; reverse: 5′-GTCCTCTTCAGAAATAAGCTTTTGTTCGGAT-3′; Integrated DNA Technologies). .. The mutagenized PCR products were further purified, re-amplified for additional 30 cycles in the absence of analog nucleotides, and combined with Sal I-HF, Nhe I-HF, and Bam HI-HI (New England BioLabs) digested pCT-CON vector. .. Pre-mixed DNA linearized vector and PCR insert (1 μg μL−1 ) was electroporated into freshly prepare EBY100 competent cells, where the full constructs are reassembled via homologous recombination .

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    New England Biolabs restriction enzyme nhei hf
    The inhibitory hairpin within the shuT RNA thermometer gradually opens as environmental temperature increases. In vitro transcribed RNA molecules were radio‐labeled at the 5′ end, and then partially digested by RNase T1, which specifically cut immediately 3′ to single‐stranded guanines. (a) Schematic of the shuT RNA thermometer with the Shine‐Dalgarno (SD) sequence highlighted with a line, the start codon in bold text and sequence from the engineered <t>NheI</t> site bracketed. All potential RNase T1 cutting sites are indicated with arrows. (b) Representative gel showing the radio‐labeled bands generated by digestion of the in vitro transcribed shuT RNA molecules. Control lanes: lane C contains RNA samples prior to the enzymatic or alkaline digestion, showing the background digestion of the experiment processes, lane L contains the sequencing ladder, and lane T1 contains the bands generated by RNase T1 digestion of the denatured template. Experimental lanes: lanes labeled 25 and <t>37°C</t> contain the bands resulting from digestion of the shuT RNA thermometer at the indicated temperature with RNase T1 at a 5‐fold dilution. (c) As a means to increase the sensitivity of the assay, it was repeated using a lesser concentration of RNase (10‐fold diluted) and the results quantified. Relative intensities of each band were normalized to that of the G27‐associated band in the same lane. G27 is predicted to be single‐stranded and thus subject to equal digestion at each temperature tested. All statistics were generated from three independent repeats. Error bars indicate one standard deviation. “*” indicates a statistically significance difference ( p ‐value ≤ .05)
    Restriction Enzyme Nhei 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/restriction enzyme nhei hf/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction enzyme nhei hf - by Bioz Stars, 2021-03
    95/100 stars
      Buy from Supplier

    Image Search Results


    The inhibitory hairpin within the shuT RNA thermometer gradually opens as environmental temperature increases. In vitro transcribed RNA molecules were radio‐labeled at the 5′ end, and then partially digested by RNase T1, which specifically cut immediately 3′ to single‐stranded guanines. (a) Schematic of the shuT RNA thermometer with the Shine‐Dalgarno (SD) sequence highlighted with a line, the start codon in bold text and sequence from the engineered NheI site bracketed. All potential RNase T1 cutting sites are indicated with arrows. (b) Representative gel showing the radio‐labeled bands generated by digestion of the in vitro transcribed shuT RNA molecules. Control lanes: lane C contains RNA samples prior to the enzymatic or alkaline digestion, showing the background digestion of the experiment processes, lane L contains the sequencing ladder, and lane T1 contains the bands generated by RNase T1 digestion of the denatured template. Experimental lanes: lanes labeled 25 and 37°C contain the bands resulting from digestion of the shuT RNA thermometer at the indicated temperature with RNase T1 at a 5‐fold dilution. (c) As a means to increase the sensitivity of the assay, it was repeated using a lesser concentration of RNase (10‐fold diluted) and the results quantified. Relative intensities of each band were normalized to that of the G27‐associated band in the same lane. G27 is predicted to be single‐stranded and thus subject to equal digestion at each temperature tested. All statistics were generated from three independent repeats. Error bars indicate one standard deviation. “*” indicates a statistically significance difference ( p ‐value ≤ .05)

    Journal: MicrobiologyOpen

    Article Title: Transcriptional and posttranscriptional regulation of Shigella shuT in response to host‐associated iron availability and temperature. Transcriptional and posttranscriptional regulation of Shigella shuT in response to host‐associated iron availability and temperature

    doi: 10.1002/mbo3.442

    Figure Lengend Snippet: The inhibitory hairpin within the shuT RNA thermometer gradually opens as environmental temperature increases. In vitro transcribed RNA molecules were radio‐labeled at the 5′ end, and then partially digested by RNase T1, which specifically cut immediately 3′ to single‐stranded guanines. (a) Schematic of the shuT RNA thermometer with the Shine‐Dalgarno (SD) sequence highlighted with a line, the start codon in bold text and sequence from the engineered NheI site bracketed. All potential RNase T1 cutting sites are indicated with arrows. (b) Representative gel showing the radio‐labeled bands generated by digestion of the in vitro transcribed shuT RNA molecules. Control lanes: lane C contains RNA samples prior to the enzymatic or alkaline digestion, showing the background digestion of the experiment processes, lane L contains the sequencing ladder, and lane T1 contains the bands generated by RNase T1 digestion of the denatured template. Experimental lanes: lanes labeled 25 and 37°C contain the bands resulting from digestion of the shuT RNA thermometer at the indicated temperature with RNase T1 at a 5‐fold dilution. (c) As a means to increase the sensitivity of the assay, it was repeated using a lesser concentration of RNase (10‐fold diluted) and the results quantified. Relative intensities of each band were normalized to that of the G27‐associated band in the same lane. G27 is predicted to be single‐stranded and thus subject to equal digestion at each temperature tested. All statistics were generated from three independent repeats. Error bars indicate one standard deviation. “*” indicates a statistically significance difference ( p ‐value ≤ .05)

    Article Snippet: Following extraction, purified pT7‐T was linearized by digestion with restriction enzyme NheI‐HF (New England Biolabs Inc.) at 37°C for 2 hr.

    Techniques: In Vitro, Labeling, Sequencing, Generated, Concentration Assay, Standard Deviation