esp3i  (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:
    Esp3I
    Description:

    Catalog Number:
    R0734
    Price:
    350
    Category:
    Restriction Enzymes
    Applications:
    DNA Manipulation
    Size:
    1500 units
    Buy from Supplier


    Structured Review

    New England Biolabs esp3i
    Esp3I

    https://www.bioz.com/result/esp3i/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    esp3i - by Bioz Stars, 2021-07
    95/100 stars

    Images

    Related Articles

    Recombinant:

    Article Title: CRTC1/MAML2 directs a PGC-1α-IGF-1 circuit that confers vulnerability to PPARγ inhibition
    Article Snippet: The pgRNA-CKB vector (Addgene #73501) ( ) was used to constitutively express the target guide RNAs. sgRNAs were cloned into the pgRNA-CKB vector using previously published methods ( ). .. Briefly, the pgRNACKB backbone was digested with Esp3I (NEB #R0734) and dephosphorylated using recombinant Shrimp Alkaline Phosphatase. .. The required sgRNAs were synthesized (Eton Biosciences, Inc) as oligonucleotides with Esp3I compatible overhangs ( ).

    Plasmid Preparation:

    Article Title: N6-methyladenosine mRNA marking promotes selective translation of regulons required for human erythropoiesis
    Article Snippet: The reaction is incubated for 4 min at 95 °C, and then 70 °C for 10 min, followed by a slow ramp to RT over a couple of hours. .. The vector is linearized with Esp3I and gel purified using the Monarch Gel Extraction Kit (NEB) or Zymoclean Gel DNA Recovery Kit (Zymo Research). .. 1 μL of annealed oligos were then ligated into the Esp3I site of 20 ng of pLL3.7-EF1a-mini with T4 ligase (NEB) for 4 h at 16 °C.

    Article Title: Knock-in and precise nucleotide substitution using near-PAMless engineered Cas9 variants in Dictyostelium discoideum
    Article Snippet: .. The annealed oligonucleotides (1.0 µl) were then ligated into an all-in-one vector (25 ng) via a Golden Gate digestion/ligation reaction using 70 U T4 DNA ligase and 1.5 U BpiI (Thermo) or Esp3I (NEB) in 4.0 µl reactions. ..

    Article Title: Fine-mapping within eQTL credible intervals by expression CROP-seq
    Article Snippet: .. The CROPseq-Guide-Puro plasmid [ ] (Addgene, Watertown MA, catalog number #86708, originally from Christoph Bock’s lab) was digested by Esp3I (NEB, R0734S). .. For each designed gRNA sequence, a pair of annealed oligos was cloned into the vector before the gRNA scaffold and after the U6 promoter.

    Article Title: Controlling gene expression in mammalian cells using multiplexed conditional guide RNAs for Cas12a
    Article Snippet: Most constructs for gRNAs or strand displacement triggers were assembled using GoldenGate cloning in combination with overlapping oligos. .. DNA oligos were purchased from IDT and annealed at a concentration of 2 μM in H2 O with 2 mM MgCl2 by cooling them from 90°C to 20°C over a period of 20 min. For the GoldenGate reactions, 3 nM of plasmid, 15 nM of each overlapping oligo pair, 1 U/μl Esp3I (NEB), 0.5 U/μl T4 Polynucleotide Kinase (NEB), and 40 U/μl T4 DNA Ligase (NEB) were mixed in T4 DNA Ligase Reaction Buffer (NEB). .. The mix was cycled between 37°C for 60s and 16°C for 90s fifteen times, kept at 37°C for 10 min, deactivated at 70°C for 10 min and chemically transformed into DH5alpha.

    Article Title: Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
    Article Snippet: .. Assembly of the MD expression constructs was conducted with 75 ng destination vector and equimolar amounts of the respective fragments, 20 U Bbs I-HF, 10 U Esp 3I, and 200 U T4-DNA ligase (NEB, Frankfurt, Germany) for 30 cycles (1 min; 16 °C; 37 °C). .. For the reference constructs, VH and VL genes were amplified incorporating SapI restriction sites and then inserted into a pTT5-derived vector utilizing CH1-CH2-CH3 or κ/λ entry vectors using GGC as described before [ , ].

    Purification:

    Article Title: N6-methyladenosine mRNA marking promotes selective translation of regulons required for human erythropoiesis
    Article Snippet: The reaction is incubated for 4 min at 95 °C, and then 70 °C for 10 min, followed by a slow ramp to RT over a couple of hours. .. The vector is linearized with Esp3I and gel purified using the Monarch Gel Extraction Kit (NEB) or Zymoclean Gel DNA Recovery Kit (Zymo Research). .. 1 μL of annealed oligos were then ligated into the Esp3I site of 20 ng of pLL3.7-EF1a-mini with T4 ligase (NEB) for 4 h at 16 °C.

    Gel Extraction:

    Article Title: N6-methyladenosine mRNA marking promotes selective translation of regulons required for human erythropoiesis
    Article Snippet: The reaction is incubated for 4 min at 95 °C, and then 70 °C for 10 min, followed by a slow ramp to RT over a couple of hours. .. The vector is linearized with Esp3I and gel purified using the Monarch Gel Extraction Kit (NEB) or Zymoclean Gel DNA Recovery Kit (Zymo Research). .. 1 μL of annealed oligos were then ligated into the Esp3I site of 20 ng of pLL3.7-EF1a-mini with T4 ligase (NEB) for 4 h at 16 °C.

    other:

    Article Title: A nanoluciferase SARS-CoV-2 for rapid neutralization testing and screening of anti-infective drugs for COVID-19
    Article Snippet: F5 and F6 fragments were obtained by digesting the plasmids with enzymes Esp3I and PvuI.

    Concentration Assay:

    Article Title: Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59
    Article Snippet: A consensus sequence for each of the cloned fragments was determined, and a consensus clone was assembled by using restriction enzymes and standard recombinant DNA techniques. .. Each of the plasmids was grown to a high concentration, isolated, and digested, or double digested, with Esp 3I, Bgl I, or Not I according to the manufacturer's directions (New England Biolabs) (Fig. ). .. The MHV-A59 A clone was digested with Mlu I, which is located in the Topo II plasmid, treated with calf intestine alkaline phosphatase, and subsequently digested with Esp 3I.

    Article Title: Controlling gene expression in mammalian cells using multiplexed conditional guide RNAs for Cas12a
    Article Snippet: Most constructs for gRNAs or strand displacement triggers were assembled using GoldenGate cloning in combination with overlapping oligos. .. DNA oligos were purchased from IDT and annealed at a concentration of 2 μM in H2 O with 2 mM MgCl2 by cooling them from 90°C to 20°C over a period of 20 min. For the GoldenGate reactions, 3 nM of plasmid, 15 nM of each overlapping oligo pair, 1 U/μl Esp3I (NEB), 0.5 U/μl T4 Polynucleotide Kinase (NEB), and 40 U/μl T4 DNA Ligase (NEB) were mixed in T4 DNA Ligase Reaction Buffer (NEB). .. The mix was cycled between 37°C for 60s and 16°C for 90s fifteen times, kept at 37°C for 10 min, deactivated at 70°C for 10 min and chemically transformed into DH5alpha.

    Isolation:

    Article Title: Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59
    Article Snippet: A consensus sequence for each of the cloned fragments was determined, and a consensus clone was assembled by using restriction enzymes and standard recombinant DNA techniques. .. Each of the plasmids was grown to a high concentration, isolated, and digested, or double digested, with Esp 3I, Bgl I, or Not I according to the manufacturer's directions (New England Biolabs) (Fig. ). .. The MHV-A59 A clone was digested with Mlu I, which is located in the Topo II plasmid, treated with calf intestine alkaline phosphatase, and subsequently digested with Esp 3I.

    End-sequence Profiling:

    Article Title: Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59
    Article Snippet: A consensus sequence for each of the cloned fragments was determined, and a consensus clone was assembled by using restriction enzymes and standard recombinant DNA techniques. .. Each of the plasmids was grown to a high concentration, isolated, and digested, or double digested, with Esp 3I, Bgl I, or Not I according to the manufacturer's directions (New England Biolabs) (Fig. ). .. The MHV-A59 A clone was digested with Mlu I, which is located in the Topo II plasmid, treated with calf intestine alkaline phosphatase, and subsequently digested with Esp 3I.

    Expressing:

    Article Title: Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
    Article Snippet: .. Assembly of the MD expression constructs was conducted with 75 ng destination vector and equimolar amounts of the respective fragments, 20 U Bbs I-HF, 10 U Esp 3I, and 200 U T4-DNA ligase (NEB, Frankfurt, Germany) for 30 cycles (1 min; 16 °C; 37 °C). .. For the reference constructs, VH and VL genes were amplified incorporating SapI restriction sites and then inserted into a pTT5-derived vector utilizing CH1-CH2-CH3 or κ/λ entry vectors using GGC as described before [ , ].

    Construct:

    Article Title: Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System
    Article Snippet: .. Assembly of the MD expression constructs was conducted with 75 ng destination vector and equimolar amounts of the respective fragments, 20 U Bbs I-HF, 10 U Esp 3I, and 200 U T4-DNA ligase (NEB, Frankfurt, Germany) for 30 cycles (1 min; 16 °C; 37 °C). .. For the reference constructs, VH and VL genes were amplified incorporating SapI restriction sites and then inserted into a pTT5-derived vector utilizing CH1-CH2-CH3 or κ/λ entry vectors using GGC as described before [ , ].

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    New England Biolabs esp 3i
    Schematic illustration of BiDi promoter system for antibody production. ( A ) The MD vector was designed to exhibit a 200-bp stuffer, flanked by <t>Esp</t> 3I restriction sites ( Esp 3I sites A and B), adjacent to a SV40 polyA signal sequence and the regions encoding for hinge-CH2-CH3, terminated by a SV40 polyA signal sequence. ( B ) VL-CL and VH-CH1 amplicons can be inserted into MD by Golden Gate cloning utilizing Esp 3I restriction sites ( Esp 3I sites A and B). The BiDi promoter can be chosen individually and is flanked by Bbs I sites ( Bbs I sites A–D), compatible with the VL and VH sequences. ( C ) Golden Gate assembly results in a fully functional and re-circularized vector, with the light chain under the control of promoter I and the heavy chain under the control of promoter II. ( D ) Schematic representation of the resulting heterotetrameric IgG1 antibody using the same colour code as for the genetic elements.
    Esp 3i, 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/esp 3i/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    esp 3i - by Bioz Stars, 2021-07
    95/100 stars
      Buy from Supplier

    Image Search Results


    Schematic illustration of BiDi promoter system for antibody production. ( A ) The MD vector was designed to exhibit a 200-bp stuffer, flanked by Esp 3I restriction sites ( Esp 3I sites A and B), adjacent to a SV40 polyA signal sequence and the regions encoding for hinge-CH2-CH3, terminated by a SV40 polyA signal sequence. ( B ) VL-CL and VH-CH1 amplicons can be inserted into MD by Golden Gate cloning utilizing Esp 3I restriction sites ( Esp 3I sites A and B). The BiDi promoter can be chosen individually and is flanked by Bbs I sites ( Bbs I sites A–D), compatible with the VL and VH sequences. ( C ) Golden Gate assembly results in a fully functional and re-circularized vector, with the light chain under the control of promoter I and the heavy chain under the control of promoter II. ( D ) Schematic representation of the resulting heterotetrameric IgG1 antibody using the same colour code as for the genetic elements.

    Journal: Antibodies

    Article Title: Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System

    doi: 10.3390/antib10020018

    Figure Lengend Snippet: Schematic illustration of BiDi promoter system for antibody production. ( A ) The MD vector was designed to exhibit a 200-bp stuffer, flanked by Esp 3I restriction sites ( Esp 3I sites A and B), adjacent to a SV40 polyA signal sequence and the regions encoding for hinge-CH2-CH3, terminated by a SV40 polyA signal sequence. ( B ) VL-CL and VH-CH1 amplicons can be inserted into MD by Golden Gate cloning utilizing Esp 3I restriction sites ( Esp 3I sites A and B). The BiDi promoter can be chosen individually and is flanked by Bbs I sites ( Bbs I sites A–D), compatible with the VL and VH sequences. ( C ) Golden Gate assembly results in a fully functional and re-circularized vector, with the light chain under the control of promoter I and the heavy chain under the control of promoter II. ( D ) Schematic representation of the resulting heterotetrameric IgG1 antibody using the same colour code as for the genetic elements.

    Article Snippet: Assembly of the MD expression constructs was conducted with 75 ng destination vector and equimolar amounts of the respective fragments, 20 U Bbs I-HF, 10 U Esp 3I, and 200 U T4-DNA ligase (NEB, Frankfurt, Germany) for 30 cycles (1 min; 16 °C; 37 °C).

    Techniques: Plasmid Preparation, Sequencing, Clone Assay, Functional Assay

    Identification of marker mutations in recombinant virus. Various icMHV-A59#1 or wild-type virus amplicons were subcloned into Topo II vectors and sequenced. (A) icMHV-A59#1 sequence across the mutated 3512 Esp 3I site; (B) icMHV-A59#1 sequence across the mutated 48 Esp 3I site and the MHV A/B junction; (C) icMHV-A59#1 sequence across the mutated 17 Esp 3I site; (D) icMHV-A59#1 sequence across the Rsr II site and MHV F/G junction; (E) wild-type MHV-A59 sequence across the same domain as shown in panel D. Overlined sequences include the mutated restriction sites and the corresponding wild-type nucleotides at these positions. In panels B and D, the bracketed sequences represent the MHV A/B and F/G junction domains, respectively. Note that all evidence of the Esp 3I site, which had been engineered into the sequence, is gone.

    Journal: Journal of Virology

    Article Title: Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59

    doi: 10.1128/JVI.76.21.11065-11078.2002

    Figure Lengend Snippet: Identification of marker mutations in recombinant virus. Various icMHV-A59#1 or wild-type virus amplicons were subcloned into Topo II vectors and sequenced. (A) icMHV-A59#1 sequence across the mutated 3512 Esp 3I site; (B) icMHV-A59#1 sequence across the mutated 48 Esp 3I site and the MHV A/B junction; (C) icMHV-A59#1 sequence across the mutated 17 Esp 3I site; (D) icMHV-A59#1 sequence across the Rsr II site and MHV F/G junction; (E) wild-type MHV-A59 sequence across the same domain as shown in panel D. Overlined sequences include the mutated restriction sites and the corresponding wild-type nucleotides at these positions. In panels B and D, the bracketed sequences represent the MHV A/B and F/G junction domains, respectively. Note that all evidence of the Esp 3I site, which had been engineered into the sequence, is gone.

    Article Snippet: Each of the plasmids was grown to a high concentration, isolated, and digested, or double digested, with Esp 3I, Bgl I, or Not I according to the manufacturer's directions (New England Biolabs) (Fig. ).

    Techniques: Marker, Recombinant, Sequencing, End-sequence Profiling

    Restriction fragment length polymorphism analysis of molecularly cloned virus. Cultures were infected with MHV-A59 or icMHV-A59#1, and intracellular RNA was isolated at 8 h p.i. With primer pairs and RT-PCR, cDNA amplicons were isolated that contained the various marker mutations that had been inserted into the component clones. The purified wild-type MHV-A59 and icMHV-A59#1 amplicons were restricted with Esp 3I or Rsr II and separated in 0.8% agarose gels. Lane 1, wild-type A59 amplicon from nucleotides 2020 to 5031, uncut; lane 2, wild-type A59 amplicon from nucleotides 2020 to 5031 digested with Esp 3I; lane 3, icMHV-A59#1 amplicon from nucleotides 2020 to 5031, uncut; lane 4, icMHV-A59#1 amplicon from nucleotides 2020 to 5031, Esp 3I digested; lane 5, wild-type A59 amplicon from nucleotides 16351 to 17875, uncut; lane 6, wild-type A59 amplicon from nucleotides 16351 to 17875 restricted with Esp 3I; lane 7, icMHV-A59#1 amplicon from nucleotides 16351 to 17875, uncut; lane 8, icMHV-A59#1 amplicon from nucleotides 16351 to 17875 restricted with Esp 3I; lane 9, wild-type A59 amplicon from nucleotides 22060 to 25416, uncut; lane 10, wild-type A59 amplicon from nucleotides 22060 to 25416 restricted with Rsr II; lane 11, icMHV-A59#1 amplicon from nucleotides 22060 to 25416, uncut; lane 12, icMHV-A59#1 amplicon from nucleotides 22060 to 25416 restricted with Rsr II; lane 13, 1-kb ladder.

    Journal: Journal of Virology

    Article Title: Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59

    doi: 10.1128/JVI.76.21.11065-11078.2002

    Figure Lengend Snippet: Restriction fragment length polymorphism analysis of molecularly cloned virus. Cultures were infected with MHV-A59 or icMHV-A59#1, and intracellular RNA was isolated at 8 h p.i. With primer pairs and RT-PCR, cDNA amplicons were isolated that contained the various marker mutations that had been inserted into the component clones. The purified wild-type MHV-A59 and icMHV-A59#1 amplicons were restricted with Esp 3I or Rsr II and separated in 0.8% agarose gels. Lane 1, wild-type A59 amplicon from nucleotides 2020 to 5031, uncut; lane 2, wild-type A59 amplicon from nucleotides 2020 to 5031 digested with Esp 3I; lane 3, icMHV-A59#1 amplicon from nucleotides 2020 to 5031, uncut; lane 4, icMHV-A59#1 amplicon from nucleotides 2020 to 5031, Esp 3I digested; lane 5, wild-type A59 amplicon from nucleotides 16351 to 17875, uncut; lane 6, wild-type A59 amplicon from nucleotides 16351 to 17875 restricted with Esp 3I; lane 7, icMHV-A59#1 amplicon from nucleotides 16351 to 17875, uncut; lane 8, icMHV-A59#1 amplicon from nucleotides 16351 to 17875 restricted with Esp 3I; lane 9, wild-type A59 amplicon from nucleotides 22060 to 25416, uncut; lane 10, wild-type A59 amplicon from nucleotides 22060 to 25416 restricted with Rsr II; lane 11, icMHV-A59#1 amplicon from nucleotides 22060 to 25416, uncut; lane 12, icMHV-A59#1 amplicon from nucleotides 22060 to 25416 restricted with Rsr II; lane 13, 1-kb ladder.

    Article Snippet: Each of the plasmids was grown to a high concentration, isolated, and digested, or double digested, with Esp 3I, Bgl I, or Not I according to the manufacturer's directions (New England Biolabs) (Fig. ).

    Techniques: Clone Assay, Infection, Isolation, Reverse Transcription Polymerase Chain Reaction, Marker, Purification, End-sequence Profiling, Amplification