agei  (New England Biolabs)


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    Name:
    AgeI
    Description:
    AgeI 1 500 units
    Catalog Number:
    r0552l
    Price:
    298
    Category:
    Restriction Enzymes
    Size:
    1 500 units
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    Structured Review

    New England Biolabs agei
    AgeI
    AgeI 1 500 units
    https://www.bioz.com/result/agei/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agei - by Bioz Stars, 2021-03
    94/100 stars

    Images

    1) Product Images from "Characterization of untranslated regions of the salmonid alphavirus 3 (SAV3) genome and construction of a SAV3 based replicon"

    Article Title: Characterization of untranslated regions of the salmonid alphavirus 3 (SAV3) genome and construction of a SAV3 based replicon

    Journal: Virology Journal

    doi: 10.1186/1743-422X-6-173

    Construction and evaluation of a SAV3 based replicon . A) A SAV3 replicon is launched from the CMV promoter (red arrow) in the pVAX1 backbone, and transcription stops at the BGH polyA signal (red box). A synthetic DNA encoding a hammerhead ribozyme was fused to the SAV3 5'-UTR and a polyadenylated tail was fused to the 3'-UTR. The ORF encoding the SAV3 structural proteins was replaced by an ORF encoding EGFP inserted between introduced AgeI and AscI sites. Position of the EcoRI site used for restriction enzyme analysis is indicated. B) Restriction enzyme analysis by digestion of pmSAV3 with EcoRI, AgeI and AscI. Lane 1: Smartladder (Eurogentec). Lane 2: pmSAV3 after triple digest with EcoRI, AgeI and AscI. Bands corresponding to the pVAX1 backbone, nsP coding sequence and EGFP coding sequence are indicated. C) Expression of EGFP in BF2 cells after transfection with pmSAV3. Both CHSE and BF2 cells facilitated successful expression of the EGFP reporter. EGFP expression became visible from day 2 p.t. in CHSE cells and 3 d.p.t. in BF2 cells.
    Figure Legend Snippet: Construction and evaluation of a SAV3 based replicon . A) A SAV3 replicon is launched from the CMV promoter (red arrow) in the pVAX1 backbone, and transcription stops at the BGH polyA signal (red box). A synthetic DNA encoding a hammerhead ribozyme was fused to the SAV3 5'-UTR and a polyadenylated tail was fused to the 3'-UTR. The ORF encoding the SAV3 structural proteins was replaced by an ORF encoding EGFP inserted between introduced AgeI and AscI sites. Position of the EcoRI site used for restriction enzyme analysis is indicated. B) Restriction enzyme analysis by digestion of pmSAV3 with EcoRI, AgeI and AscI. Lane 1: Smartladder (Eurogentec). Lane 2: pmSAV3 after triple digest with EcoRI, AgeI and AscI. Bands corresponding to the pVAX1 backbone, nsP coding sequence and EGFP coding sequence are indicated. C) Expression of EGFP in BF2 cells after transfection with pmSAV3. Both CHSE and BF2 cells facilitated successful expression of the EGFP reporter. EGFP expression became visible from day 2 p.t. in CHSE cells and 3 d.p.t. in BF2 cells.

    Techniques Used: Sequencing, Expressing, Transfection

    2) Product Images from "Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System"

    Article Title: Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System

    Journal: Journal of Virology

    doi: 10.1128/JVI.02956-12

    Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal
    Figure Legend Snippet: Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal

    Techniques Used:

    3) Product Images from "Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System"

    Article Title: Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System

    Journal: Journal of Virology

    doi: 10.1128/JVI.02956-12

    Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal
    Figure Legend Snippet: Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal

    Techniques Used:

    Related Articles

    Sequencing:

    Article Title: Critical factors for the bulk adhesion of engineered elastomeric proteins
    Article Snippet: The elastin-based proteins ELP[KEY4 -24], ELP[KEY4 -48], ELP[KEY4 -96] and ELP[K3 Y3 -48] were constructed with a cloning scheme modified from one previously developed by our laboratory [ ]. .. The new scheme used AgeI and AvaI restriction enzymes (New England Biolabs, Ipswich, MA) to achieve seamless repeats of the elastin-like sequence. .. Standard molecular cloning techniques were used throughout [ ].

    Article Title: Characterization of untranslated regions of the salmonid alphavirus 3 (SAV3) genome and construction of a SAV3 based replicon
    Article Snippet: .. The authenticity of the plasmid construction was verified by EcoRI, AgeI and AscI (New England Biolabs) digestion (Fig. ) and by sequencing as previously described [ ]. .. This information indicated that eight substitutions were present in the RC coding region compared to the nucleotide sequence of passage 20 of the parental strain SAVH20/03 (Table ).

    Isolation:

    Article Title: The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli
    Article Snippet: The RT-PCR product was analyzed on 10% PAA gels in TBE buffer, and purified using ‘PCR clean up kit’ (Qiagen, Hilden, Germany). .. For AgeI restriction analysis the isolated RT-PCR product was incubated with 4 units of AgeI-HF (New England Biolabs) for 30 min at 37°C. .. The restriction fragments were determined using PAA gel-electrophoresis and visualized by EtBr-staining.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli
    Article Snippet: The RT-PCR product was analyzed on 10% PAA gels in TBE buffer, and purified using ‘PCR clean up kit’ (Qiagen, Hilden, Germany). .. For AgeI restriction analysis the isolated RT-PCR product was incubated with 4 units of AgeI-HF (New England Biolabs) for 30 min at 37°C. .. The restriction fragments were determined using PAA gel-electrophoresis and visualized by EtBr-staining.

    Incubation:

    Article Title: The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli
    Article Snippet: The RT-PCR product was analyzed on 10% PAA gels in TBE buffer, and purified using ‘PCR clean up kit’ (Qiagen, Hilden, Germany). .. For AgeI restriction analysis the isolated RT-PCR product was incubated with 4 units of AgeI-HF (New England Biolabs) for 30 min at 37°C. .. The restriction fragments were determined using PAA gel-electrophoresis and visualized by EtBr-staining.

    Plasmid Preparation:

    Article Title: Opposing Actions of Hippocampus TNFα Receptors on Limbic Seizure Susceptibility
    Article Snippet: Coding sequences were PCR amplified (Phusion DNA polymerase, New England Biolabs, Ipswich, MA) using oligomers (Integrated DNA Technologies, Coralville, Iowa) with AgeI-forward and Not1-reverse overhangs incorporated. .. Amplicons were digested with AgeI and NotI restriction enzymes (New England Biolabs) and ligated into pTR2/CBA-GFP, acquired from the UNC Gene Therapy Center Vector Core. ..

    Article Title: CORALINA: a universal method for the generation of gRNA libraries for CRISPR-based screening
    Article Snippet: For lentiviral vector construction, the vector pLKO.1 (Addgene plasmid 10878) was modified to insert the gRNA promoter and scaffolding sequence from pgRNA1. .. The vector was first digested with EcoRI (NEB) and AgeI (NEB). .. Next, the desired sequences were amplified from pgRNA1 using primers gRNA-PLKO-F (5′-TTTCTTGGGTAGTTTGCAGTTTT) and gRNA-PLKO-R (5′-ccatttgtctcgaggtcgag-TACCTCGAGCGGCCCAAGC) and inserted into PLKO.1.

    Article Title: Characterization of untranslated regions of the salmonid alphavirus 3 (SAV3) genome and construction of a SAV3 based replicon
    Article Snippet: .. The authenticity of the plasmid construction was verified by EcoRI, AgeI and AscI (New England Biolabs) digestion (Fig. ) and by sequencing as previously described [ ]. .. This information indicated that eight substitutions were present in the RC coding region compared to the nucleotide sequence of passage 20 of the parental strain SAVH20/03 (Table ).

    Purification:

    Article Title: Clonal analysis of human anti-V3 monoclonal antibodies selected by a V3 tetramer
    Article Snippet: .. Purified PCR products were digested with the respective restriction enzymes AgeI, SalI, BsiWI, and XhoI (NEB), ligated into expression vectors and transformed into Competent E.coli DH10B bacteria (Invitrogen). .. Ampicillin resistant colonies were selected and screened by PCR for inserts of the expected size (650 bp for Ig γ , 700 bp for Ig κ and 590 bp for Ig λ ) and then sequenced to confirm the identity with the original PCR amplicons.

    Polymerase Chain Reaction:

    Article Title: Clonal analysis of human anti-V3 monoclonal antibodies selected by a V3 tetramer
    Article Snippet: .. Purified PCR products were digested with the respective restriction enzymes AgeI, SalI, BsiWI, and XhoI (NEB), ligated into expression vectors and transformed into Competent E.coli DH10B bacteria (Invitrogen). .. Ampicillin resistant colonies were selected and screened by PCR for inserts of the expected size (650 bp for Ig γ , 700 bp for Ig κ and 590 bp for Ig λ ) and then sequenced to confirm the identity with the original PCR amplicons.

    Expressing:

    Article Title: Clonal analysis of human anti-V3 monoclonal antibodies selected by a V3 tetramer
    Article Snippet: .. Purified PCR products were digested with the respective restriction enzymes AgeI, SalI, BsiWI, and XhoI (NEB), ligated into expression vectors and transformed into Competent E.coli DH10B bacteria (Invitrogen). .. Ampicillin resistant colonies were selected and screened by PCR for inserts of the expected size (650 bp for Ig γ , 700 bp for Ig κ and 590 bp for Ig λ ) and then sequenced to confirm the identity with the original PCR amplicons.

    Transformation Assay:

    Article Title: Clonal analysis of human anti-V3 monoclonal antibodies selected by a V3 tetramer
    Article Snippet: .. Purified PCR products were digested with the respective restriction enzymes AgeI, SalI, BsiWI, and XhoI (NEB), ligated into expression vectors and transformed into Competent E.coli DH10B bacteria (Invitrogen). .. Ampicillin resistant colonies were selected and screened by PCR for inserts of the expected size (650 bp for Ig γ , 700 bp for Ig κ and 590 bp for Ig λ ) and then sequenced to confirm the identity with the original PCR amplicons.

    Ligation:

    Article Title: Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System
    Article Snippet: The endonuclease reaction mixtures were purified, and the C92 and P98 amplicons were ligated to each other using T4 DNA ligase (NEB). .. The ligation reaction mixture was then digested with AgeI (NEB) and ligated to the AgeI-digested del-C92-TOPO “subclone” ( ). .. The ligation reaction mixture was transformed into E. coli , and the transformants were screened via colony PCR using the Topo-seq-F and Topo-seq-R primers ( ).

    Article Title: Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System
    Article Snippet: The endonuclease reaction mixtures were purified, and the C92 and P98 amplicons were ligated to each other using T4 DNA ligase (NEB). .. The ligation reaction mixture was digested with AgeI (NEB) and ligated to AgeI-digested del-C92-TOPO “subclone” ( ). .. The ligation reaction mixture was transformed into E. coli , and colonies were screened by colony PCR using the Topo-seq-F and Topo-seq-R primers ( ).

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  • 94
    New England Biolabs agei
    Construction and evaluation of a SAV3 based replicon . A) A SAV3 replicon is launched from the CMV promoter (red arrow) in the pVAX1 backbone, and transcription stops at the BGH polyA signal (red box). A synthetic DNA encoding a hammerhead ribozyme was fused to the SAV3 5'-UTR and a polyadenylated tail was fused to the 3'-UTR. The ORF encoding the SAV3 structural proteins was replaced by an ORF encoding EGFP inserted between introduced <t>AgeI</t> and AscI sites. Position of the <t>EcoRI</t> site used for restriction enzyme analysis is indicated. B) Restriction enzyme analysis by digestion of pmSAV3 with EcoRI, AgeI and AscI. Lane 1: Smartladder (Eurogentec). Lane 2: pmSAV3 after triple digest with EcoRI, AgeI and AscI. Bands corresponding to the pVAX1 backbone, nsP coding sequence and EGFP coding sequence are indicated. C) Expression of EGFP in BF2 cells after transfection with pmSAV3. Both CHSE and BF2 cells facilitated successful expression of the EGFP reporter. EGFP expression became visible from day 2 p.t. in CHSE cells and 3 d.p.t. in BF2 cells.
    Agei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/agei/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    agei - by Bioz Stars, 2021-03
    94/100 stars
      Buy from Supplier

    Image Search Results


    Construction and evaluation of a SAV3 based replicon . A) A SAV3 replicon is launched from the CMV promoter (red arrow) in the pVAX1 backbone, and transcription stops at the BGH polyA signal (red box). A synthetic DNA encoding a hammerhead ribozyme was fused to the SAV3 5'-UTR and a polyadenylated tail was fused to the 3'-UTR. The ORF encoding the SAV3 structural proteins was replaced by an ORF encoding EGFP inserted between introduced AgeI and AscI sites. Position of the EcoRI site used for restriction enzyme analysis is indicated. B) Restriction enzyme analysis by digestion of pmSAV3 with EcoRI, AgeI and AscI. Lane 1: Smartladder (Eurogentec). Lane 2: pmSAV3 after triple digest with EcoRI, AgeI and AscI. Bands corresponding to the pVAX1 backbone, nsP coding sequence and EGFP coding sequence are indicated. C) Expression of EGFP in BF2 cells after transfection with pmSAV3. Both CHSE and BF2 cells facilitated successful expression of the EGFP reporter. EGFP expression became visible from day 2 p.t. in CHSE cells and 3 d.p.t. in BF2 cells.

    Journal: Virology Journal

    Article Title: Characterization of untranslated regions of the salmonid alphavirus 3 (SAV3) genome and construction of a SAV3 based replicon

    doi: 10.1186/1743-422X-6-173

    Figure Lengend Snippet: Construction and evaluation of a SAV3 based replicon . A) A SAV3 replicon is launched from the CMV promoter (red arrow) in the pVAX1 backbone, and transcription stops at the BGH polyA signal (red box). A synthetic DNA encoding a hammerhead ribozyme was fused to the SAV3 5'-UTR and a polyadenylated tail was fused to the 3'-UTR. The ORF encoding the SAV3 structural proteins was replaced by an ORF encoding EGFP inserted between introduced AgeI and AscI sites. Position of the EcoRI site used for restriction enzyme analysis is indicated. B) Restriction enzyme analysis by digestion of pmSAV3 with EcoRI, AgeI and AscI. Lane 1: Smartladder (Eurogentec). Lane 2: pmSAV3 after triple digest with EcoRI, AgeI and AscI. Bands corresponding to the pVAX1 backbone, nsP coding sequence and EGFP coding sequence are indicated. C) Expression of EGFP in BF2 cells after transfection with pmSAV3. Both CHSE and BF2 cells facilitated successful expression of the EGFP reporter. EGFP expression became visible from day 2 p.t. in CHSE cells and 3 d.p.t. in BF2 cells.

    Article Snippet: The authenticity of the plasmid construction was verified by EcoRI, AgeI and AscI (New England Biolabs) digestion (Fig. ) and by sequencing as previously described [ ].

    Techniques: Sequencing, Expressing, Transfection

    RtcB re-ligates RNA43 to purified 70S Δ43 ribosomes in vitro . ( A ) Schematic showing the in vitro ribosome repair assay (see text for details). ( B ) RNA43 AgeI used in the assay. Nucleotides changed in helix 45 and the binding site for primers Y12 (gray) and H17 (green) are indicated. ( C ) RT-PCR performed on RNA purified from 70S Δ43 ribosomes incubated in the absence of RtcB and RNA43 AgeI (lane 1), in the presence of either RNA43 AgeI (lane 2) or RtcB (lane 3), or both (lane 4) employing primer pairs specific for the central region of 16S rRNA (S7/X15; panel a), the 3΄end of 16S rRNA (S7/Y12; panel b), and specific for the AU nucleotide change present in 16S AgeI rRNA upon successful ligation (S7/H17; panel c). ( D ) AgeI restriction analysis and ( E ), sequencing of the RT-PCR product obtained with primer pair S7/H17 (shown in C, panel c, lane 4). Sequencing analyses from position 1509–1517 of the 16S rRNA of (a) rrsB , and (b) the RT-PCR product are shown. Nucleotides changed in RNA43 AgeI are underlined.

    Journal: Nucleic Acids Research

    Article Title: The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli

    doi: 10.1093/nar/gkw1018

    Figure Lengend Snippet: RtcB re-ligates RNA43 to purified 70S Δ43 ribosomes in vitro . ( A ) Schematic showing the in vitro ribosome repair assay (see text for details). ( B ) RNA43 AgeI used in the assay. Nucleotides changed in helix 45 and the binding site for primers Y12 (gray) and H17 (green) are indicated. ( C ) RT-PCR performed on RNA purified from 70S Δ43 ribosomes incubated in the absence of RtcB and RNA43 AgeI (lane 1), in the presence of either RNA43 AgeI (lane 2) or RtcB (lane 3), or both (lane 4) employing primer pairs specific for the central region of 16S rRNA (S7/X15; panel a), the 3΄end of 16S rRNA (S7/Y12; panel b), and specific for the AU nucleotide change present in 16S AgeI rRNA upon successful ligation (S7/H17; panel c). ( D ) AgeI restriction analysis and ( E ), sequencing of the RT-PCR product obtained with primer pair S7/H17 (shown in C, panel c, lane 4). Sequencing analyses from position 1509–1517 of the 16S rRNA of (a) rrsB , and (b) the RT-PCR product are shown. Nucleotides changed in RNA43 AgeI are underlined.

    Article Snippet: For AgeI restriction analysis the isolated RT-PCR product was incubated with 4 units of AgeI-HF (New England Biolabs) for 30 min at 37°C.

    Techniques: Purification, In Vitro, Binding Assay, Reverse Transcription Polymerase Chain Reaction, Incubation, Ligation, Sequencing

    Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal

    Journal: Journal of Virology

    Article Title: Insights into a Viral Lytic Pathway from an Archaeal Virus-Host System

    doi: 10.1128/JVI.02956-12

    Figure Lengend Snippet: Schematic overview of the construction of the C92/P98 chimeric genes. (A) Construction of N-terminal C92 + C-terminal P98 for replication within STIV. P1, c92 + AgeI-F; P2, c92 + BsrGI-R; P3, p98 + BsrGI-F; P4, p98 + AgeI-R. (B) Construction of N-terminal

    Article Snippet: The ligation reaction mixture was digested with AgeI (NEB) and ligated to AgeI-digested del-C92-TOPO “subclone” ( ).

    Techniques: