esp3i  (New England Biolabs)


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    Name:
    Esp3I
    Description:
    Esp3I 1 500 units
    Catalog Number:
    r0734l
    Price:
    343
    Category:
    Restriction Enzymes
    Size:
    1 500 units
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    New England Biolabs esp3i
    Esp3I
    Esp3I 1 500 units
    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, 2021-03
    95/100 stars

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    Related Articles

    Plasmid Preparation:

    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: A One-Step Process for the Construction of Phage Display scFv and VHH Libraries.
    Article Snippet: In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes.

    Article Title: Multimodal Analysis of Composition and Spatial Architecture in Human Squamous Cell Carcinoma
    Article Snippet: The amplified oligo pool was run on a 2% agarose gel and purified using the Zymoclean Gel Recovery kit. .. 16ng of the oligo pool was then assembled with 446ng Esp3I digested pLentiguide vector in a Gibson Assembly reaction (NEB). .. The reaction was transformed into Stellar competent cells and grown overnight in liquid culture with ampicillin selection.

    Article Title: Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
    Article Snippet: For generating landing pad vectors from expression vectors without the correct overhangs, an oligonucleotide stuffer was used to complete the overhangs. .. TA-splitHalo BFP plasmids were made in 10 μL reactions comprising 20 fmol Kanamycin ColE1 digested backbone, 40 fmol TA-splitHalo fusion expression vectors, 40 fmol PGK-mTagBFP2 expression vector, 10x T4 Ligase Buffer (NEB B0202S), Esp31 (NEB R0734S/L), and T7 DNA Ligase (NEB M0318S/L) with the same cycling conditions as the part vectors. .. Transfection of HeLa cells in 8-well chamber flasks for TA-splitHalo Architecture BenchmarkingFor transfections, HeLa cells were seeded in an 8-well chamber flask at 20k cells per well in 225 μL DMEM +1% Penicillin/Streptomycin (P/S) 10% Fetal Bovine Serum (FBS).

    Article Title: Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
    Article Snippet: Expression vectors were generated in 10 μL reactions containing 20 fmol CDS backbone, 40 fmol of each part insert, 10x T4 Ligase Buffer, BsaI-HF v2.0 (NEB R3733S/L), and T7 DNA Ligase with the same cycling conditions as the part vectors. .. Landing pad (LP) vectors were generated similarly to part vectors in 10 μL reactions with 20 fmol MTK landing pad entry backbone (Addgene #123932), 40 fmol of each expression vector plasmids, 10x T4 Ligase Buffer (NEB B0202S), Esp31 (NEB R0734S/L), and T7 DNA Ligase (M0318S/L) with the same cycling conditions as the part vectors. .. For generating landing pad vectors from expression vectors without the correct overhangs, an oligonucleotide stuffer was used to complete the overhangs.

    Article Title: A bispecific immunotweezer prevents soluble PrP oligomers and abolishes prion toxicity
    Article Snippet: Reaction mix was heated for 4 min at 95°C on a heating block ThermoStat (Eppendorf), then the heating block was turned off and the reaction was allowed to proceed for 30 min on the block and was then put at 4°C. .. Golden Gate assembly [ ] was used in order to clone the double-stranded DNA Oligomers into the MLM3636 plasmid, using the following reaction: 150 ng MLM3636 plasmid 1 μL double stranded oligomer ligation mix 2 μL T4 ligase buffer (New England Biolabs) 13.25 μL ddH2 O 1 μL Esp3I (New England Biolabs) 1 μL T4 ligase (New England Biolabs) This reaction was put on a thermocycler using the following conditions: 37°C, 5 minutes 16°C, 10 minutes (10 cycles with these two steps) 37°C, 15 minutes 80°C, 5 minutes The ligated plasmid MLM3636(sgRNAmPrnp ) was subsequently transformed into DH5α chemically competent E . coli cells (Invitrogen) and plasmid purification was undertaken using Plasmid Maxi Kit (Qiagen). .. CAD5 cells were co-transfected using the MLM3636(sgRNAmPrnp ) plasmid and the hCas9 plasmid (hCas9 was a gift from George Church, Addgene plasmid # 41815, [ ]) dissolved in Lipofectamine 2000 (Invitrogen).

    Polymerase Chain Reaction:

    Article Title: A One-Step Process for the Construction of Phage Display scFv and VHH Libraries.
    Article Snippet: In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes.

    Purification:

    Article Title: A One-Step Process for the Construction of Phage Display scFv and VHH Libraries.
    Article Snippet: In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes.

    Article Title: A bispecific immunotweezer prevents soluble PrP oligomers and abolishes prion toxicity
    Article Snippet: Reaction mix was heated for 4 min at 95°C on a heating block ThermoStat (Eppendorf), then the heating block was turned off and the reaction was allowed to proceed for 30 min on the block and was then put at 4°C. .. Golden Gate assembly [ ] was used in order to clone the double-stranded DNA Oligomers into the MLM3636 plasmid, using the following reaction: 150 ng MLM3636 plasmid 1 μL double stranded oligomer ligation mix 2 μL T4 ligase buffer (New England Biolabs) 13.25 μL ddH2 O 1 μL Esp3I (New England Biolabs) 1 μL T4 ligase (New England Biolabs) This reaction was put on a thermocycler using the following conditions: 37°C, 5 minutes 16°C, 10 minutes (10 cycles with these two steps) 37°C, 15 minutes 80°C, 5 minutes The ligated plasmid MLM3636(sgRNAmPrnp ) was subsequently transformed into DH5α chemically competent E . coli cells (Invitrogen) and plasmid purification was undertaken using Plasmid Maxi Kit (Qiagen). .. CAD5 cells were co-transfected using the MLM3636(sgRNAmPrnp ) plasmid and the hCas9 plasmid (hCas9 was a gift from George Church, Addgene plasmid # 41815, [ ]) dissolved in Lipofectamine 2000 (Invitrogen).

    Amplification:

    Article Title: A One-Step Process for the Construction of Phage Display scFv and VHH Libraries.
    Article Snippet: In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes.

    Sequencing:

    Article Title: A One-Step Process for the Construction of Phage Display scFv and VHH Libraries.
    Article Snippet: In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes. .. In this work we present a one-step cloning approach for the establishment of antibody phage display libraries relying on type IIs restriction enzymes.

    Expressing:

    Article Title: Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
    Article Snippet: For generating landing pad vectors from expression vectors without the correct overhangs, an oligonucleotide stuffer was used to complete the overhangs. .. TA-splitHalo BFP plasmids were made in 10 μL reactions comprising 20 fmol Kanamycin ColE1 digested backbone, 40 fmol TA-splitHalo fusion expression vectors, 40 fmol PGK-mTagBFP2 expression vector, 10x T4 Ligase Buffer (NEB B0202S), Esp31 (NEB R0734S/L), and T7 DNA Ligase (NEB M0318S/L) with the same cycling conditions as the part vectors. .. Transfection of HeLa cells in 8-well chamber flasks for TA-splitHalo Architecture BenchmarkingFor transfections, HeLa cells were seeded in an 8-well chamber flask at 20k cells per well in 225 μL DMEM +1% Penicillin/Streptomycin (P/S) 10% Fetal Bovine Serum (FBS).

    Article Title: Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
    Article Snippet: Expression vectors were generated in 10 μL reactions containing 20 fmol CDS backbone, 40 fmol of each part insert, 10x T4 Ligase Buffer, BsaI-HF v2.0 (NEB R3733S/L), and T7 DNA Ligase with the same cycling conditions as the part vectors. .. Landing pad (LP) vectors were generated similarly to part vectors in 10 μL reactions with 20 fmol MTK landing pad entry backbone (Addgene #123932), 40 fmol of each expression vector plasmids, 10x T4 Ligase Buffer (NEB B0202S), Esp31 (NEB R0734S/L), and T7 DNA Ligase (M0318S/L) with the same cycling conditions as the part vectors. .. For generating landing pad vectors from expression vectors without the correct overhangs, an oligonucleotide stuffer was used to complete the overhangs.

    Generated:

    Article Title: Versatile labeling and detection of endogenous proteins using tag-assisted split enzyme complementation
    Article Snippet: Expression vectors were generated in 10 μL reactions containing 20 fmol CDS backbone, 40 fmol of each part insert, 10x T4 Ligase Buffer, BsaI-HF v2.0 (NEB R3733S/L), and T7 DNA Ligase with the same cycling conditions as the part vectors. .. Landing pad (LP) vectors were generated similarly to part vectors in 10 μL reactions with 20 fmol MTK landing pad entry backbone (Addgene #123932), 40 fmol of each expression vector plasmids, 10x T4 Ligase Buffer (NEB B0202S), Esp31 (NEB R0734S/L), and T7 DNA Ligase (M0318S/L) with the same cycling conditions as the part vectors. .. For generating landing pad vectors from expression vectors without the correct overhangs, an oligonucleotide stuffer was used to complete the overhangs.

    Ligation:

    Article Title: A bispecific immunotweezer prevents soluble PrP oligomers and abolishes prion toxicity
    Article Snippet: Reaction mix was heated for 4 min at 95°C on a heating block ThermoStat (Eppendorf), then the heating block was turned off and the reaction was allowed to proceed for 30 min on the block and was then put at 4°C. .. Golden Gate assembly [ ] was used in order to clone the double-stranded DNA Oligomers into the MLM3636 plasmid, using the following reaction: 150 ng MLM3636 plasmid 1 μL double stranded oligomer ligation mix 2 μL T4 ligase buffer (New England Biolabs) 13.25 μL ddH2 O 1 μL Esp3I (New England Biolabs) 1 μL T4 ligase (New England Biolabs) This reaction was put on a thermocycler using the following conditions: 37°C, 5 minutes 16°C, 10 minutes (10 cycles with these two steps) 37°C, 15 minutes 80°C, 5 minutes The ligated plasmid MLM3636(sgRNAmPrnp ) was subsequently transformed into DH5α chemically competent E . coli cells (Invitrogen) and plasmid purification was undertaken using Plasmid Maxi Kit (Qiagen). .. CAD5 cells were co-transfected using the MLM3636(sgRNAmPrnp ) plasmid and the hCas9 plasmid (hCas9 was a gift from George Church, Addgene plasmid # 41815, [ ]) dissolved in Lipofectamine 2000 (Invitrogen).

    Transformation Assay:

    Article Title: A bispecific immunotweezer prevents soluble PrP oligomers and abolishes prion toxicity
    Article Snippet: Reaction mix was heated for 4 min at 95°C on a heating block ThermoStat (Eppendorf), then the heating block was turned off and the reaction was allowed to proceed for 30 min on the block and was then put at 4°C. .. Golden Gate assembly [ ] was used in order to clone the double-stranded DNA Oligomers into the MLM3636 plasmid, using the following reaction: 150 ng MLM3636 plasmid 1 μL double stranded oligomer ligation mix 2 μL T4 ligase buffer (New England Biolabs) 13.25 μL ddH2 O 1 μL Esp3I (New England Biolabs) 1 μL T4 ligase (New England Biolabs) This reaction was put on a thermocycler using the following conditions: 37°C, 5 minutes 16°C, 10 minutes (10 cycles with these two steps) 37°C, 15 minutes 80°C, 5 minutes The ligated plasmid MLM3636(sgRNAmPrnp ) was subsequently transformed into DH5α chemically competent E . coli cells (Invitrogen) and plasmid purification was undertaken using Plasmid Maxi Kit (Qiagen). .. CAD5 cells were co-transfected using the MLM3636(sgRNAmPrnp ) plasmid and the hCas9 plasmid (hCas9 was a gift from George Church, Addgene plasmid # 41815, [ ]) dissolved in Lipofectamine 2000 (Invitrogen).

    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.

    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.

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    New England Biolabs esp 3i
    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 <t>Esp</t> 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.
    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, 2021-03
    95/100 stars
      Buy from Supplier

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    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