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agei restriction site oka300  (New England Biolabs)


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    New England Biolabs agei restriction site oka300
    Agei Restriction Site Oka300, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 870 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/agei+restriction+site/AgeI/bio_rxiv__2025__10__07__680855-202-11-16
    Average 97 stars, based on 870 article reviews
    agei restriction site oka300 - by Bioz Stars, 2026-09
    97/100 stars

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    Related Articles

    Synthesized:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Introduce:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Clone Assay:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Polymerase Chain Reaction:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Article Title: Mapping of Functional Subdomains in the Terminal Protein Domain of Hepatitis B Virus Polymerase
    Article Snippet: .. To facilitate PCR-mediated manipulations, an AgeI restriction site was introduced using the New England BioLabs Q5 site-directed mutagenesis kit. ..

    Article Title: Nodal patterning without Lefty inhibitory feedback is functional but fragile
    Article Snippet: .. Optimal PCR con- ditions: Taq polymerase, 25 cycles, 57 ̊C annealing temperature. lefty2 wild type genotyping forward primer: cattttgaccacagcgat lefty2 mutant genotyping forward primer: gttcattttgaccactcac The common reverse primer was identical to lefty2 reverse primer described above. squint: The squintcz35 allele has a ~ 1.9 kb insertion in exon 1, and was detected as in (Feldman et al., 1998). cyclops: The cyclopsm294 mutation destroys an AgeI restriction site, and was detected as in (Sampath et al., 1998). lefty and squint expression constructs To generate mRNA from all constructs, plasmids were linearized with NotI-HF endonuclease (NEB) and purified using a Qiagen PCR clean-up kit. .. Capped mRNA was generated from linearized plasmid using an SP6 mMessage mMachine kit (Ambion) and purified with a Qiagen RNeasy kit.

    Article Title: Mapping of Functional Subdomains in the Terminal Protein Domain of Hepatitis B Virus Polymerase
    Article Snippet: .. To facilitate PCR 158 on N ovem ber 18, 2016 by U N IV O F T O R O N T O http://jvi.asm .org/ D ow nloaded from 9 mediated manipulations, an AgeI restriction site was introduced using the New England Biolabs 159 Q5 site-directed mutagenesis kit. ..

    Amplification:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Article Title: Two Type VI Secretion Systems in Vibrio coralliilyticus RE22Sm exhibit differential target specificity for bacteria prey and oyster larvae
    Article Snippet: .. The Km resistance gene was amplified from the TOPO2.1 vector (Invitrogen) and inserted into pDM5 via the Gibson Assembly Reaction at the AgeI restriction site. pDM5 was linearized at the SacI restriction enzyme site, using SacI-HF (New England Biolabs), within the multicloning region (MCR) for all mutation destined Gibson Assemblies. .. The ligation mixture was introduced into E. coli Sm10 (containing λpir) by electroporation with the BioRad Gene Pulser II in a 2 mm cuvette (2.5 kV; 25 μF; 200 Ω).

    Expressing:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Article Title: Nodal patterning without Lefty inhibitory feedback is functional but fragile
    Article Snippet: .. Optimal PCR con- ditions: Taq polymerase, 25 cycles, 57 ̊C annealing temperature. lefty2 wild type genotyping forward primer: cattttgaccacagcgat lefty2 mutant genotyping forward primer: gttcattttgaccactcac The common reverse primer was identical to lefty2 reverse primer described above. squint: The squintcz35 allele has a ~ 1.9 kb insertion in exon 1, and was detected as in (Feldman et al., 1998). cyclops: The cyclopsm294 mutation destroys an AgeI restriction site, and was detected as in (Sampath et al., 1998). lefty and squint expression constructs To generate mRNA from all constructs, plasmids were linearized with NotI-HF endonuclease (NEB) and purified using a Qiagen PCR clean-up kit. .. Capped mRNA was generated from linearized plasmid using an SP6 mMessage mMachine kit (Ambion) and purified with a Qiagen RNeasy kit.

    Plasmid Preparation:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Sequencing:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Cloning:

    Article Title: Real-time reporter systems for monitoring base editing
    Article Snippet: .. A gBlock, synthesized by Integrated DNA Technologies (IDT) to introduce a CMV promoter with a 3′ AgeI restriction site, was cloned into the nef open reading frame using BamHI and KpnI restriction sites. mCherry was PCR amplified using Phusion high-fidelity DNA polymerase (NEB) from a pcDNA3.1 expression plasmid with primers that introduced a 3′ T2A self-cleaving peptide sequence (primer sequences provided in TABLE 1) and cloned into a CloneJET PCR cloning vector (Thermo Fisher). eGFP was PCR amplified from a pcDNA3.1 expression plasmid with primers (TABLE 1) introducing scrambled nucleotide sequences at the 5′ and 3′ ends of the gene that retained the wild-type protein sequence. ..

    Mutagenesis:

    Article Title: Mapping of Functional Subdomains in the Terminal Protein Domain of Hepatitis B Virus Polymerase
    Article Snippet: .. To facilitate PCR-mediated manipulations, an AgeI restriction site was introduced using the New England BioLabs Q5 site-directed mutagenesis kit. ..

    Article Title: Two Type VI Secretion Systems in Vibrio coralliilyticus RE22Sm exhibit differential target specificity for bacteria prey and oyster larvae
    Article Snippet: .. The Km resistance gene was amplified from the TOPO2.1 vector (Invitrogen) and inserted into pDM5 via the Gibson Assembly Reaction at the AgeI restriction site. pDM5 was linearized at the SacI restriction enzyme site, using SacI-HF (New England Biolabs), within the multicloning region (MCR) for all mutation destined Gibson Assemblies. .. The ligation mixture was introduced into E. coli Sm10 (containing λpir) by electroporation with the BioRad Gene Pulser II in a 2 mm cuvette (2.5 kV; 25 μF; 200 Ω).

    Article Title: Nodal patterning without Lefty inhibitory feedback is functional but fragile
    Article Snippet: .. Optimal PCR con- ditions: Taq polymerase, 25 cycles, 57 ̊C annealing temperature. lefty2 wild type genotyping forward primer: cattttgaccacagcgat lefty2 mutant genotyping forward primer: gttcattttgaccactcac The common reverse primer was identical to lefty2 reverse primer described above. squint: The squintcz35 allele has a ~ 1.9 kb insertion in exon 1, and was detected as in (Feldman et al., 1998). cyclops: The cyclopsm294 mutation destroys an AgeI restriction site, and was detected as in (Sampath et al., 1998). lefty and squint expression constructs To generate mRNA from all constructs, plasmids were linearized with NotI-HF endonuclease (NEB) and purified using a Qiagen PCR clean-up kit. .. Capped mRNA was generated from linearized plasmid using an SP6 mMessage mMachine kit (Ambion) and purified with a Qiagen RNeasy kit.

    Article Title: Mapping of Functional Subdomains in the Terminal Protein Domain of Hepatitis B Virus Polymerase
    Article Snippet: .. To facilitate PCR 158 on N ovem ber 18, 2016 by U N IV O F T O R O N T O http://jvi.asm .org/ D ow nloaded from 9 mediated manipulations, an AgeI restriction site was introduced using the New England Biolabs 159 Q5 site-directed mutagenesis kit. ..

    Article Title: Functional and structural characterization of mouse Factor H-related B protein unveils a novel dimerization domain shared by FHR-B and FH
    Article Snippet: The two C-terminal SCR domains of mouse FHR-B, FHR-C (UniPort E9Q8B5) and FHR-E (UniProt Q61406) were generated with GeneArt Gene Synthesis (Thermo Scientific) in the pMA-T vector flanked by the AgeI (5’) and MluI (3’) restriction sites and cloned into the pMA-T vector. .. For the generation of the FHR-1 1-3 ::mFH 19-20 hybrid protein, a new AgeI restriction site, which resulted in synonymous codons, was generated between SCRs 3-4 of FHR-1 and SCRs 18-19 of mFH by site-directed mutagenesis (New England Biolabs, #E0554S). ..

    Construct:

    Article Title: Nodal patterning without Lefty inhibitory feedback is functional but fragile
    Article Snippet: .. Optimal PCR con- ditions: Taq polymerase, 25 cycles, 57 ̊C annealing temperature. lefty2 wild type genotyping forward primer: cattttgaccacagcgat lefty2 mutant genotyping forward primer: gttcattttgaccactcac The common reverse primer was identical to lefty2 reverse primer described above. squint: The squintcz35 allele has a ~ 1.9 kb insertion in exon 1, and was detected as in (Feldman et al., 1998). cyclops: The cyclopsm294 mutation destroys an AgeI restriction site, and was detected as in (Sampath et al., 1998). lefty and squint expression constructs To generate mRNA from all constructs, plasmids were linearized with NotI-HF endonuclease (NEB) and purified using a Qiagen PCR clean-up kit. .. Capped mRNA was generated from linearized plasmid using an SP6 mMessage mMachine kit (Ambion) and purified with a Qiagen RNeasy kit.

    Purification:

    Article Title: Nodal patterning without Lefty inhibitory feedback is functional but fragile
    Article Snippet: .. Optimal PCR con- ditions: Taq polymerase, 25 cycles, 57 ̊C annealing temperature. lefty2 wild type genotyping forward primer: cattttgaccacagcgat lefty2 mutant genotyping forward primer: gttcattttgaccactcac The common reverse primer was identical to lefty2 reverse primer described above. squint: The squintcz35 allele has a ~ 1.9 kb insertion in exon 1, and was detected as in (Feldman et al., 1998). cyclops: The cyclopsm294 mutation destroys an AgeI restriction site, and was detected as in (Sampath et al., 1998). lefty and squint expression constructs To generate mRNA from all constructs, plasmids were linearized with NotI-HF endonuclease (NEB) and purified using a Qiagen PCR clean-up kit. .. Capped mRNA was generated from linearized plasmid using an SP6 mMessage mMachine kit (Ambion) and purified with a Qiagen RNeasy kit.

    Generated:

    Article Title: Functional and structural characterization of mouse Factor H-related B protein unveils a novel dimerization domain shared by FHR-B and FH
    Article Snippet: The two C-terminal SCR domains of mouse FHR-B, FHR-C (UniPort E9Q8B5) and FHR-E (UniProt Q61406) were generated with GeneArt Gene Synthesis (Thermo Scientific) in the pMA-T vector flanked by the AgeI (5’) and MluI (3’) restriction sites and cloned into the pMA-T vector. .. For the generation of the FHR-1 1-3 ::mFH 19-20 hybrid protein, a new AgeI restriction site, which resulted in synonymous codons, was generated between SCRs 3-4 of FHR-1 and SCRs 18-19 of mFH by site-directed mutagenesis (New England Biolabs, #E0554S). ..



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