sapi  (New England Biolabs)


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  • 94
    Name:
    SapI
    Description:
    SapI 1 250 units
    Catalog Number:
    R0569L
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    1 250 units
    Buy from Supplier


    Structured Review

    New England Biolabs sapi
    SapI
    SapI 1 250 units
    https://www.bioz.com/result/sapi/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi - by Bioz Stars, 2021-05
    94/100 stars

    Images

    1) Product Images from "Development of the SapI/AarI Incision Mediated Plasmid Editing Method"

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method

    Journal: Journal of molecular biology

    doi: 10.1016/j.jmb.2018.03.030

    Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.
    Figure Legend Snippet: Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.

    Techniques Used: Sequencing, Mutagenesis

    2) Product Images from "A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology"

    Article Title: A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2013.00339

    Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.
    Figure Legend Snippet: Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.

    Techniques Used: Plasmid Preparation, Clone Assay, Produced

    3) Product Images from "CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo"

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo

    Journal: Current protocols in molecular biology

    doi: 10.1002/cpmb.46

    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.
    Figure Legend Snippet: Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Techniques Used: Plasmid Preparation, Clone Assay, Construct

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Changing the topology of protein backbone: the effect of backbone cyclization on the structure and dynamics of a SH3 domain
    Article Snippet: .. The PCR amplified SH3 domain was purified, digested simultaneously with NdeI and SapI and then ligated into a NdeI-SapI-treated plasmid pTXB-1 (New England Biolabs). .. The resulting plasmid pTXB-1-SH3lin−wt was shown to be free of mutations in the c-Crk SH3-encoding region by DNA sequencing.

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method
    Article Snippet: All PCR reactions were performed using the Phusion High-Fidelity DNA Polymerase (NEB), using the manufacturer’s recommended specifications. .. After amplification, PCR products were first digested with DpnI (NEB) to eliminate the parental plasmid, cleaned up over columns (Qiagen), and then digested with either the SapI (NEB) or AarI (Thermo) restriction enzyme, following the manufacturer’s specifications. .. After digestion, DNA products were cleaned up again as before, and 50μg of the DNA was self-ligated using T4 DNA ligase (NEB).

    Amplification:

    Article Title: Changing the topology of protein backbone: the effect of backbone cyclization on the structure and dynamics of a SH3 domain
    Article Snippet: .. The PCR amplified SH3 domain was purified, digested simultaneously with NdeI and SapI and then ligated into a NdeI-SapI-treated plasmid pTXB-1 (New England Biolabs). .. The resulting plasmid pTXB-1-SH3lin−wt was shown to be free of mutations in the c-Crk SH3-encoding region by DNA sequencing.

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: .. To clone in the additional functional components between the guide and donor, we amplified and cloned in the sgtail and terminator sequences following the same Golden Gate cloning method as described above, but this time using SapI (NEB R0569S) and T4 ligase. .. The resulting Golden Gate reactions were then PCR purified and electroporated into 5-alpha Electrocompetent E.coli cells to create a final guide+donor library.

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method
    Article Snippet: All PCR reactions were performed using the Phusion High-Fidelity DNA Polymerase (NEB), using the manufacturer’s recommended specifications. .. After amplification, PCR products were first digested with DpnI (NEB) to eliminate the parental plasmid, cleaned up over columns (Qiagen), and then digested with either the SapI (NEB) or AarI (Thermo) restriction enzyme, following the manufacturer’s specifications. .. After digestion, DNA products were cleaned up again as before, and 50μg of the DNA was self-ligated using T4 DNA ligase (NEB).

    Purification:

    Article Title: Changing the topology of protein backbone: the effect of backbone cyclization on the structure and dynamics of a SH3 domain
    Article Snippet: .. The PCR amplified SH3 domain was purified, digested simultaneously with NdeI and SapI and then ligated into a NdeI-SapI-treated plasmid pTXB-1 (New England Biolabs). .. The resulting plasmid pTXB-1-SH3lin−wt was shown to be free of mutations in the c-Crk SH3-encoding region by DNA sequencing.

    Plasmid Preparation:

    Article Title: Changing the topology of protein backbone: the effect of backbone cyclization on the structure and dynamics of a SH3 domain
    Article Snippet: .. The PCR amplified SH3 domain was purified, digested simultaneously with NdeI and SapI and then ligated into a NdeI-SapI-treated plasmid pTXB-1 (New England Biolabs). .. The resulting plasmid pTXB-1-SH3lin−wt was shown to be free of mutations in the c-Crk SH3-encoding region by DNA sequencing.

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method
    Article Snippet: All PCR reactions were performed using the Phusion High-Fidelity DNA Polymerase (NEB), using the manufacturer’s recommended specifications. .. After amplification, PCR products were first digested with DpnI (NEB) to eliminate the parental plasmid, cleaned up over columns (Qiagen), and then digested with either the SapI (NEB) or AarI (Thermo) restriction enzyme, following the manufacturer’s specifications. .. After digestion, DNA products were cleaned up again as before, and 50μg of the DNA was self-ligated using T4 DNA ligase (NEB).

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo
    Article Snippet: The protocol is written for cloning of a single gRNA, but the protocol can be scaled to conveniently clone multiple gRNAs in parallel. .. AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019). ..

    Functional Assay:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: .. To clone in the additional functional components between the guide and donor, we amplified and cloned in the sgtail and terminator sequences following the same Golden Gate cloning method as described above, but this time using SapI (NEB R0569S) and T4 ligase. .. The resulting Golden Gate reactions were then PCR purified and electroporated into 5-alpha Electrocompetent E.coli cells to create a final guide+donor library.

    Clone Assay:

    Article Title: High-throughput creation and functional profiling of DNA sequence variant libraries using CRISPR/Cas9 in yeast
    Article Snippet: .. To clone in the additional functional components between the guide and donor, we amplified and cloned in the sgtail and terminator sequences following the same Golden Gate cloning method as described above, but this time using SapI (NEB R0569S) and T4 ligase. .. The resulting Golden Gate reactions were then PCR purified and electroporated into 5-alpha Electrocompetent E.coli cells to create a final guide+donor library.

    other:

    Article Title: Replication-coupled DNA Interstrand Crosslink repair in Xenopus egg extracts
    Article Snippet: HincII and SapI restriction enzymes (NEB).

    Nucleic Acid Electrophoresis:

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo
    Article Snippet: The protocol is written for cloning of a single gRNA, but the protocol can be scaled to conveniently clone multiple gRNAs in parallel. .. AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019). ..

    Gel Purification:

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo
    Article Snippet: The protocol is written for cloning of a single gRNA, but the protocol can be scaled to conveniently clone multiple gRNAs in parallel. .. AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019). ..

    Ligation:

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo
    Article Snippet: The protocol is written for cloning of a single gRNA, but the protocol can be scaled to conveniently clone multiple gRNAs in parallel. .. AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019). ..

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  • 94
    New England Biolabs sapi enzyme
    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed <t>oligos</t> corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and <t>SapI</t> sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.
    Sapi Enzyme, 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/sapi enzyme/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sapi enzyme - by Bioz Stars, 2021-05
    94/100 stars
      Buy from Supplier

    Image Search Results


    Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Journal: Current protocols in molecular biology

    Article Title: CASAAV: a CRISPR based platform for rapid dissection of gene function in vivo

    doi: 10.1002/cpmb.46

    Figure Lengend Snippet: Insertion of gRNA sequences into AAV vector gRNAs are inserted into AAV plasmid containing U6 promoters and cardiac troponin T (cTNT) promoter-driven Cre recombinase. Annealed oligos corresponding to the targeting portions of gRNA1 and gRNA2 are cloned between AarI and SapI sites, respectively. The entire construct is contained within AAV inverted terminal repeat sequences, allowing for packaging into AAV capsids. Examples of gRNAs designed against Gata4 are shown.

    Article Snippet: AAV-U6gRNA1-U6gRNA2-TnT-Cre Vector (Addgene #87682) gRNA sequences from Basic Protocol 1/DNA oligos Thermocycler AarI enzyme and buffer (ThermoFisher ER1581) SapI enzyme and buffer (NEB R0569L) 37°C incubator 37°C shaking incubator Agarose (GeneMate E-3120-500) 50× TAE (Boston BioProducts BM250) Ethidium bromide (Sigma E7637) DNA loading dye (NEB B7024S) Gel electrophoresis chamber UV transilluminator (365 nm) Gel purification kit (Invitrogen K210012) Quick ligation kit (NEB M2200L) Chemically competent cells such as ThermoFisher MAX Efficiency Stbl2 Competent Cells (Cat# 10268019).

    Techniques: Plasmid Preparation, Clone Assay, Construct

    Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.

    Journal: Journal of molecular biology

    Article Title: Development of the SapI/AarI Incision Mediated Plasmid Editing Method

    doi: 10.1016/j.jmb.2018.03.030

    Figure Lengend Snippet: Primer design for introducing point mutations using AarI and SapI. (A) Recognition sites and cutting patterns for the SapI and AarI restriction enzymes. (B) Hypothetical sequence that will be targeted for mutagenesis. (C) Primers that can be used to mutate the sequence shown in (B) with the SIMPLE method, using the SapI enzyme.

    Article Snippet: After amplification, PCR products were first digested with DpnI (NEB) to eliminate the parental plasmid, cleaned up over columns (Qiagen), and then digested with either the SapI (NEB) or AarI (Thermo) restriction enzyme, following the manufacturer’s specifications.

    Techniques: Sequencing, Mutagenesis

    Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.

    Journal: Frontiers in Plant Science

    Article Title: A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology

    doi: 10.3389/fpls.2013.00339

    Figure Lengend Snippet: Golden Gate assembly of an insert into the destination vector. (A) Recognition sequences for the Type IIS restriction endonucleases BsaI and SapI . 5′ overhang sequences used for annealing fragments are shown in bold magenta lettering. (B) Example of a Golden Gate-compatible destination vector. Note the orientation of the BsaI sites cause excision of the lacZ α gene. (C) Example of a Golden Gate compatible vector containing a gene of interest (GOI) that will be released after digestion with BsaI . (D) A typical Golden Gate Cloning reaction would involve mixing the destination vector and insert storage vector together into one tube at equal molar ratio with BsaI and T4 DNA ligase. The final vector produced would lack BsaI recognition sequences and be resistant to digestion.

    Article Snippet: BsaI (R0535L) , SapI (R0569M, 10,000 units/ml), T4 DNA ligase (M0202M) were obtained from New England Biolabs.

    Techniques: Plasmid Preparation, Clone Assay, Produced

    Examination of dual incisions during ICL repair. HincII digested repair intermediates are separated on a denaturing gel and visualized using Southern blotting (land 5–14) (Subheading 3.4). Undigested and HincII and/or SapI digested pControl and

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Replication-coupled DNA Interstrand Crosslink repair in Xenopus egg extracts

    doi: 10.1007/978-1-61779-998-3_16

    Figure Lengend Snippet: Examination of dual incisions during ICL repair. HincII digested repair intermediates are separated on a denaturing gel and visualized using Southern blotting (land 5–14) (Subheading 3.4). Undigested and HincII and/or SapI digested pControl and

    Article Snippet: HincII and SapI restriction enzymes (NEB).

    Techniques: Southern Blot

    Determination of ICL repair efficiency. (a) Analysis of HincII (lane 1–10) and HincII/SapI (lane 11–20) digested repair intermediates on a native agarose gel (Subheading 3.5). (b) Calculated repair efficiency plotted against time.

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Replication-coupled DNA Interstrand Crosslink repair in Xenopus egg extracts

    doi: 10.1007/978-1-61779-998-3_16

    Figure Lengend Snippet: Determination of ICL repair efficiency. (a) Analysis of HincII (lane 1–10) and HincII/SapI (lane 11–20) digested repair intermediates on a native agarose gel (Subheading 3.5). (b) Calculated repair efficiency plotted against time.

    Article Snippet: HincII and SapI restriction enzymes (NEB).

    Techniques: Agarose Gel Electrophoresis