esp3i sites  (Thermo Fisher)


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  • 99
    Name:
    Esp3I BsmBI 10 U µL
    Description:
    5 C G T C T C N1↓ 3 3 G C A G A G N5↑ 5 Thermo Scientific Esp3I BsmBI restriction enzyme recognizes CGTCTC 1 5 sites and cuts best at 37°C in Tango DTT buffer See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Note Also available as a FastDigest enzyme for rapid DNA digestion Isoschizomers BsmBI Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that sent in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote The enzyme requires DTT Freshly made DTT should be added to the reaction buffer Esp3I cleaves downstream of its recognition site and can generate any desired 4 base 5 overhangs This feature is useful for PCR product cloning For methylation sensitivity refer to product specifications
    Catalog Number:
    er0451
    Price:
    None
    Applications:
    Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
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    Structured Review

    Thermo Fisher esp3i sites
    Golden Gate assembly of custom TAL effector and TALEN constructs using module, array, last repeat and backbone plasmids. By using the type IIS restriction endonucleases BsaI and <t>Esp3I,</t> modules containing the desired RVDs can be released with unique cohesive ends for ordered, single-reaction assembly into array plasmids in a first step, and those arrays subsequently released and assembled in order in a second step into a backbone plasmid to create full length constructs with custom repeat arrays (see text for details). NLS, nuclear localization signal(s); AD, transcriptional activation domain; tet , tetracycline resistance; spec , spectinomycin resistance; amp , ampicillin resistance; attL1 and attL2, recombination sites for Gateway cloning; B, BamHI, and S, SphI, useful for subcloning custom repeat arrays. Unique restriction enzyme sites flanking the coding sequences, useful for subcloning the entire constructs into other vectors, are not shown but can be found in the sequence files ( Supplementary Data ).
    5 C G T C T C N1↓ 3 3 G C A G A G N5↑ 5 Thermo Scientific Esp3I BsmBI restriction enzyme recognizes CGTCTC 1 5 sites and cuts best at 37°C in Tango DTT buffer See Reaction Conditions for Restriction Enzymes for a table of enzyme activity conditions for double digestion and heat inactivation for this and other restriction enzymes Note Also available as a FastDigest enzyme for rapid DNA digestion Isoschizomers BsmBI Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes optimized to work in one of the buffers of the Five Buffer System In addition the universal Tango buffer is provided for convenience in double digestions All of the enzymes exhibit 100 activity in the recommended buffer and reaction conditions To ensure consistent performance Thermo Scientific restriction enzyme reaction buffers contain premixed BSA which enhances the stability of many enzymes and binds contaminants that sent in DNA preparations Features• Superior quality stringent quality control and industry leading manufacturing process• Convenient color coded Five Buffer System• Includes universal Tango buffer for double digestions• BSA premixed in reaction buffers• Wide selection of restriction endonuclease specificitiesApplications• Molecular cloning• Restriction site mapping• Genotyping• Southern blotting• Restriction fragment length polymorphism RFLP • SNPNote The enzyme requires DTT Freshly made DTT should be added to the reaction buffer Esp3I cleaves downstream of its recognition site and can generate any desired 4 base 5 overhangs This feature is useful for PCR product cloning For methylation sensitivity refer to product specifications
    https://www.bioz.com/result/esp3i sites/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    esp3i sites - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting"

    Article Title: Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkr218

    Golden Gate assembly of custom TAL effector and TALEN constructs using module, array, last repeat and backbone plasmids. By using the type IIS restriction endonucleases BsaI and Esp3I, modules containing the desired RVDs can be released with unique cohesive ends for ordered, single-reaction assembly into array plasmids in a first step, and those arrays subsequently released and assembled in order in a second step into a backbone plasmid to create full length constructs with custom repeat arrays (see text for details). NLS, nuclear localization signal(s); AD, transcriptional activation domain; tet , tetracycline resistance; spec , spectinomycin resistance; amp , ampicillin resistance; attL1 and attL2, recombination sites for Gateway cloning; B, BamHI, and S, SphI, useful for subcloning custom repeat arrays. Unique restriction enzyme sites flanking the coding sequences, useful for subcloning the entire constructs into other vectors, are not shown but can be found in the sequence files ( Supplementary Data ).
    Figure Legend Snippet: Golden Gate assembly of custom TAL effector and TALEN constructs using module, array, last repeat and backbone plasmids. By using the type IIS restriction endonucleases BsaI and Esp3I, modules containing the desired RVDs can be released with unique cohesive ends for ordered, single-reaction assembly into array plasmids in a first step, and those arrays subsequently released and assembled in order in a second step into a backbone plasmid to create full length constructs with custom repeat arrays (see text for details). NLS, nuclear localization signal(s); AD, transcriptional activation domain; tet , tetracycline resistance; spec , spectinomycin resistance; amp , ampicillin resistance; attL1 and attL2, recombination sites for Gateway cloning; B, BamHI, and S, SphI, useful for subcloning custom repeat arrays. Unique restriction enzyme sites flanking the coding sequences, useful for subcloning the entire constructs into other vectors, are not shown but can be found in the sequence files ( Supplementary Data ).

    Techniques Used: Construct, Plasmid Preparation, Activation Assay, Clone Assay, Subcloning, Sequencing

    Related Articles

    shRNA:

    Article Title: Malocclusion Generates Anxiety-Like Behavior Through a Putative Lateral Habenula–Mesencephalic Trigeminal Nucleus Pathway
    Article Snippet: .. The pAAV-U6-RNAi-eGFP vector contained two Esp3I (Thermo Fisher Scientific) recognition sites downstream of the U6 promoter for shRNA adaption. .. The six resulting VGLUT2 shRNA vectors (containing sequences expressing enhanced green fluorescent protein, EGFP) were co-transfected with reporter vectors (containing sequences expressing VGLUT2 and mCherry) into HEK293 cells.

    Construct:

    Article Title: Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting
    Article Snippet: .. Finally, into this plasmid, the XbaI–SacI fragment of pTAL3 containing the TALEN backbone construct was introduced at the corresponding sites. pTAL1 was created by replacing the SphI fragment of tal1C in pCS691 with the corresponding SphI fragment of pTAL3, containing the lacZ gene and the Esp3I sites and flanking sequences for accepting final arrays. pCS691 is a derivative of Gateway entry vector pENTR-D (Invitrogen) containing between the attL sites, the complete tal1c gene preceded by both Kozak and Shine–Dalgarno consensus sequences for efficient translation in eukaryotic or bacterial cells, respectively. .. In pCS691, the kanamycin resistance gene of pENTR-D is replaced by the BspHI fragment of pBlueScript SK(-) (Stratagene) for ampicillin resistance.

    Purification:

    Article Title: Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria
    Article Snippet: .. We performed the Golden Gate assembly reaction with 1,000 ng of part4 plasmid (pHLL214 or pHLL215) and 40 ng of the barcode PCR product using BsmBI (Esp3I from Thermo Fisher) and T4 DNA ligase under the following cycling conditions: 10 cycles, with 1 cycle consisting of 5 min at 37°C and 10 min at 16°C, followed by a final digestion step at 37°C for 30 min. We then purified the Golden Gate assembly reaction with the DNA Clean & Concentrator kit (Zymo Research) and digested the DNA overnight with BsmBI. ..

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: .. The resulting dsDNA is then ligated into the BsmBI-digested pPapi vector using Golden Gate assembly: 5 μL Tango Buffer (ThermoFisher) 5 μL DTT (stored at -80°C and used once, 10 mM stock) 5 μL ATP (stored at -80°C and used once, 10 mM stock) 500 ng pPapi vector, pre-digested with Esp3I or BsmBI, gel-extracted, and isopropanol-precipitation purified 100 ng dual sgRNA dsDNA insert 1 μL Esp3I (ThermoFisher ER0452) 1 μL T7 ligase (Enzymatics, 3,000 Units / μL L6020L) Up to 50 μL water Cycle 100x (overnight): 5 minutes at 37°C, 5 minutes at 20°C. ..

    Incubation:

    Article Title: Rapid single step subcloning procedure by combined action of type II and type IIs endonucleases with ligase
    Article Snippet: .. We combined entry vector, destination vector, Esp3I (Fermentas), NheI (Fermentas) and T4 ligase (Promega) in a buffer allowing all three enzyme actions (**) and incubated at room temperature for 1 hour (Figure ). .. We transformed 2 μl of the resulting solution into DH5α chemically competent E. coli (Invitrogen) and plated on two plates either containing kanamycin or ampicillin.

    Polymerase Chain Reaction:

    Article Title: Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria
    Article Snippet: .. We performed the Golden Gate assembly reaction with 1,000 ng of part4 plasmid (pHLL214 or pHLL215) and 40 ng of the barcode PCR product using BsmBI (Esp3I from Thermo Fisher) and T4 DNA ligase under the following cycling conditions: 10 cycles, with 1 cycle consisting of 5 min at 37°C and 10 min at 16°C, followed by a final digestion step at 37°C for 30 min. We then purified the Golden Gate assembly reaction with the DNA Clean & Concentrator kit (Zymo Research) and digested the DNA overnight with BsmBI. ..

    Plasmid Preparation:

    Article Title: Rapid single step subcloning procedure by combined action of type II and type IIs endonucleases with ligase
    Article Snippet: .. We combined entry vector, destination vector, Esp3I (Fermentas), NheI (Fermentas) and T4 ligase (Promega) in a buffer allowing all three enzyme actions (**) and incubated at room temperature for 1 hour (Figure ). .. We transformed 2 μl of the resulting solution into DH5α chemically competent E. coli (Invitrogen) and plated on two plates either containing kanamycin or ampicillin.

    Article Title: Antisense probing of dynamic RNA structures.
    Article Snippet: .. RNA regulation is influenced by the dynamic changes in conformational accessibility on the transcript. .. RNA regulation is influenced by the dynamic changes in conformational accessibility on the transcript.

    Article Title: Magic Pools: Parallel Assessment of Transposon Delivery Vectors in Bacteria
    Article Snippet: .. We performed the Golden Gate assembly reaction with 1,000 ng of part4 plasmid (pHLL214 or pHLL215) and 40 ng of the barcode PCR product using BsmBI (Esp3I from Thermo Fisher) and T4 DNA ligase under the following cycling conditions: 10 cycles, with 1 cycle consisting of 5 min at 37°C and 10 min at 16°C, followed by a final digestion step at 37°C for 30 min. We then purified the Golden Gate assembly reaction with the DNA Clean & Concentrator kit (Zymo Research) and digested the DNA overnight with BsmBI. ..

    Article Title: Malocclusion Generates Anxiety-Like Behavior Through a Putative Lateral Habenula–Mesencephalic Trigeminal Nucleus Pathway
    Article Snippet: .. The pAAV-U6-RNAi-eGFP vector contained two Esp3I (Thermo Fisher Scientific) recognition sites downstream of the U6 promoter for shRNA adaption. .. The six resulting VGLUT2 shRNA vectors (containing sequences expressing enhanced green fluorescent protein, EGFP) were co-transfected with reporter vectors (containing sequences expressing VGLUT2 and mCherry) into HEK293 cells.

    Article Title: Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting
    Article Snippet: .. Finally, into this plasmid, the XbaI–SacI fragment of pTAL3 containing the TALEN backbone construct was introduced at the corresponding sites. pTAL1 was created by replacing the SphI fragment of tal1C in pCS691 with the corresponding SphI fragment of pTAL3, containing the lacZ gene and the Esp3I sites and flanking sequences for accepting final arrays. pCS691 is a derivative of Gateway entry vector pENTR-D (Invitrogen) containing between the attL sites, the complete tal1c gene preceded by both Kozak and Shine–Dalgarno consensus sequences for efficient translation in eukaryotic or bacterial cells, respectively. .. In pCS691, the kanamycin resistance gene of pENTR-D is replaced by the BspHI fragment of pBlueScript SK(-) (Stratagene) for ampicillin resistance.

    Article Title: Uncoupling of sgRNAs from their associated barcodes during PCR amplification of combinatorial CRISPR screens
    Article Snippet: .. The resulting dsDNA is then ligated into the BsmBI-digested pPapi vector using Golden Gate assembly: 5 μL Tango Buffer (ThermoFisher) 5 μL DTT (stored at -80°C and used once, 10 mM stock) 5 μL ATP (stored at -80°C and used once, 10 mM stock) 500 ng pPapi vector, pre-digested with Esp3I or BsmBI, gel-extracted, and isopropanol-precipitation purified 100 ng dual sgRNA dsDNA insert 1 μL Esp3I (ThermoFisher ER0452) 1 μL T7 ligase (Enzymatics, 3,000 Units / μL L6020L) Up to 50 μL water Cycle 100x (overnight): 5 minutes at 37°C, 5 minutes at 20°C. ..

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  • 99
    Thermo Fisher esp 3i sites
    Esp 3i Sites, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/esp 3i sites/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    esp 3i sites - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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