saci sites  (Thermo Fisher)


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    Name:
    SacI 10 U µL
    Description:
    5 G A G C T ↓C 3 3 C ↑T C G A G 5 Thermo Scientific SacI restriction enzyme recognizes GAGCT C sites and cuts best at 37°C in its own unique 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 Eco53kI EcoICRI Psp124BI SstI 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 may be present 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 SacI is sensitive to cytosine methylation at GAGmCTC but not GAGCTmC and insensitive to adenine methylation at GmAGCTC AluI methyltransferase AGmCT can be used to block SacI Supercoiled plasmids may require up to 5 fold more SacI for complete digestion than linear DNA SacI is inhibited by common clinical anticoagulants found in some preparation of anticoagulated peripheral blood and bone marrow Levels of EDTA and ACD citric acid sodium citrate dextrose in standard sample preparation have been shown to inhibit SacI Three times the normal concentration for heparin is required to inhibit SacI J E Coad et al Inhibition of restriction endonucleases by common clinical anticoagulants Anal Biochem 205 368 369 1992 For methylation sensitivity refer to product specifications
    Catalog Number:
    er1131
    Price:
    None
    Applications:
    Cloning|Restriction Enzyme Cloning
    Category:
    Proteins Enzymes Peptides
    Buy from Supplier


    Structured Review

    Thermo Fisher saci sites
    5 G A G C T ↓C 3 3 C ↑T C G A G 5 Thermo Scientific SacI restriction enzyme recognizes GAGCT C sites and cuts best at 37°C in its own unique 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 Eco53kI EcoICRI Psp124BI SstI 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 may be present 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 SacI is sensitive to cytosine methylation at GAGmCTC but not GAGCTmC and insensitive to adenine methylation at GmAGCTC AluI methyltransferase AGmCT can be used to block SacI Supercoiled plasmids may require up to 5 fold more SacI for complete digestion than linear DNA SacI is inhibited by common clinical anticoagulants found in some preparation of anticoagulated peripheral blood and bone marrow Levels of EDTA and ACD citric acid sodium citrate dextrose in standard sample preparation have been shown to inhibit SacI Three times the normal concentration for heparin is required to inhibit SacI J E Coad et al Inhibition of restriction endonucleases by common clinical anticoagulants Anal Biochem 205 368 369 1992 For methylation sensitivity refer to product specifications
    https://www.bioz.com/result/saci sites/product/Thermo Fisher
    Average 95 stars, based on 30 article reviews
    Price from $9.99 to $1999.99
    saci sites - by Bioz Stars, 2020-09
    95/100 stars

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    Related Articles

    Agarose Gel Electrophoresis:

    Article Title: Fluorescence-Based Bacterial Bioreporter for Specific Detection of Methyl Halide Emissions in the Environment
    Article Snippet: .. The resulting PCR fragment was purified after agarose gel electrophoresis using the Geneclean Turbo kit (MP Biomedicals), digested overnight with the enzymes SacI and KpnI (Fermentas), and purified again by using the same kit. .. The digested PCR fragment was ligated for 24 h at 14°C with KpnI- and SacI-digested promoter probe plasmid pLM.syfp2 (see Fig. S1 in the supplemental material), which features a promotorless gene for YFP downstream of its multiple-cloning site and a kanamycin resistance gene , and transformed into One Shot TOP10 chemically competent cells (Invitrogen) according to the manufacturer's instructions.

    Synthesized:

    Article Title: Carbon dioxide fixation by Calvin-Cycle enzymes improves ethanol yield in yeast
    Article Snippet: .. Rubisco form II gene cbbM from T. denitrificans [ ] flanked by KpnI and SacI sites was codon optimized [ ] (accession number: KC699554), synthesized at GeneArt (Life Technologies Europe BV), and ligated into pPCR-Script. .. The cbbM -containing fragment was ligated into the BamHI and SacI restricted vector pGPD_426 [ ] creating plasmid pBTWW002.

    Spectrophotometry:

    Article Title: Entericidin Is Required for a Probiotic Treatment ( Enterobacter sp. Strain C6-6) To Protect Trout from Cold-Water Disease Challenge
    Article Snippet: .. The purified PCR template and the suicide vector pDM4 were both digested using the restriction enzymes SacI and XhoI for 3 h at 37°C, purified again, quantified by using a NanoDrop ND-1000 spectrophotometer (Thermo Scientific), and ligated using T4 DNA ligase (New England BioLabs). .. The ligated product was then transformed into electrocompetent E. coli S17 λ pir ( ) and selected on LB plates with chloramphenicol.

    Purification:

    Article Title: Entericidin Is Required for a Probiotic Treatment ( Enterobacter sp. Strain C6-6) To Protect Trout from Cold-Water Disease Challenge
    Article Snippet: .. The purified PCR template and the suicide vector pDM4 were both digested using the restriction enzymes SacI and XhoI for 3 h at 37°C, purified again, quantified by using a NanoDrop ND-1000 spectrophotometer (Thermo Scientific), and ligated using T4 DNA ligase (New England BioLabs). .. The ligated product was then transformed into electrocompetent E. coli S17 λ pir ( ) and selected on LB plates with chloramphenicol.

    Article Title: Fluorescence-Based Bacterial Bioreporter for Specific Detection of Methyl Halide Emissions in the Environment
    Article Snippet: .. The resulting PCR fragment was purified after agarose gel electrophoresis using the Geneclean Turbo kit (MP Biomedicals), digested overnight with the enzymes SacI and KpnI (Fermentas), and purified again by using the same kit. .. The digested PCR fragment was ligated for 24 h at 14°C with KpnI- and SacI-digested promoter probe plasmid pLM.syfp2 (see Fig. S1 in the supplemental material), which features a promotorless gene for YFP downstream of its multiple-cloning site and a kanamycin resistance gene , and transformed into One Shot TOP10 chemically competent cells (Invitrogen) according to the manufacturer's instructions.

    Electrophoresis:

    Article Title: Efficiency of Base Excision Repair of Oxidative DNA Damage and Its Impact on the Risk of Colorectal Cancer in the Polish Population
    Article Snippet: .. A 450 pb length DNA fragment was excised from plasmid by 1 U of SacI enzyme (ThermoScientific, Rochester, USA) incubated at 37°C for 1 h. To allow the repaired and unrepaired fractions to be differentiated, the remaining unrepaired AP site was treated with 1 U of AP-recognizing endonuclease IV for 1 h. All samples were run on 8% urea-containing polyacrylamide gel for 3 h in 120 V. The accurate electrophoresis was preceded by 1 h preelectrophoresis with loading buffer. ..

    Polymerase Chain Reaction:

    Article Title: Entericidin Is Required for a Probiotic Treatment ( Enterobacter sp. Strain C6-6) To Protect Trout from Cold-Water Disease Challenge
    Article Snippet: .. The purified PCR template and the suicide vector pDM4 were both digested using the restriction enzymes SacI and XhoI for 3 h at 37°C, purified again, quantified by using a NanoDrop ND-1000 spectrophotometer (Thermo Scientific), and ligated using T4 DNA ligase (New England BioLabs). .. The ligated product was then transformed into electrocompetent E. coli S17 λ pir ( ) and selected on LB plates with chloramphenicol.

    Article Title: Fluorescence-Based Bacterial Bioreporter for Specific Detection of Methyl Halide Emissions in the Environment
    Article Snippet: .. The resulting PCR fragment was purified after agarose gel electrophoresis using the Geneclean Turbo kit (MP Biomedicals), digested overnight with the enzymes SacI and KpnI (Fermentas), and purified again by using the same kit. .. The digested PCR fragment was ligated for 24 h at 14°C with KpnI- and SacI-digested promoter probe plasmid pLM.syfp2 (see Fig. S1 in the supplemental material), which features a promotorless gene for YFP downstream of its multiple-cloning site and a kanamycin resistance gene , and transformed into One Shot TOP10 chemically competent cells (Invitrogen) according to the manufacturer's instructions.

    Incubation:

    Article Title: Efficiency of Base Excision Repair of Oxidative DNA Damage and Its Impact on the Risk of Colorectal Cancer in the Polish Population
    Article Snippet: .. A 450 pb length DNA fragment was excised from plasmid by 1 U of SacI enzyme (ThermoScientific, Rochester, USA) incubated at 37°C for 1 h. To allow the repaired and unrepaired fractions to be differentiated, the remaining unrepaired AP site was treated with 1 U of AP-recognizing endonuclease IV for 1 h. All samples were run on 8% urea-containing polyacrylamide gel for 3 h in 120 V. The accurate electrophoresis was preceded by 1 h preelectrophoresis with loading buffer. ..

    Modification:

    Article Title: Cd36, a class B scavenger receptor, functions as a monomer to bind acetylated and oxidized low-density lipoproteins
    Article Snippet: .. The modified cDNA was excised from the plasmid using restriction enzymes SacI and AgeI and subcloned into similarly digested pIZ/V5-His (Invitrogen). .. This fused, in frame, the coding sequences for the 6-histidine tag of the cDNA with the 6-histidine tag of the pIZ/V5-His vector, thus encoding a protein of full-length Cd36 with a carboxy-terminal tag of GHHHHHHTGHHHHHH.

    Transformation Assay:

    Article Title: Lectin activity of the nucleocytoplasmic EUL protein from Arabidopsis thaliana
    Article Snippet: .. Plasmid DNA from transformed E. coli was linearized using the SacI restriction enzyme (Fermentas, Sankt Leon-Rot, Germany) overnight at 37°C. .. Approximately 10 μg of the linearized DNA was transformed into Pichia strain KM-71H cells by electroporation (Bio-Rad, Hercules, CA, USA) with following pulse settings: 25 μF, 1.5 kV, 200 Ω .

    Plasmid Preparation:

    Article Title: Efficiency of Base Excision Repair of Oxidative DNA Damage and Its Impact on the Risk of Colorectal Cancer in the Polish Population
    Article Snippet: .. A 450 pb length DNA fragment was excised from plasmid by 1 U of SacI enzyme (ThermoScientific, Rochester, USA) incubated at 37°C for 1 h. To allow the repaired and unrepaired fractions to be differentiated, the remaining unrepaired AP site was treated with 1 U of AP-recognizing endonuclease IV for 1 h. All samples were run on 8% urea-containing polyacrylamide gel for 3 h in 120 V. The accurate electrophoresis was preceded by 1 h preelectrophoresis with loading buffer. ..

    Article Title: Lectin activity of the nucleocytoplasmic EUL protein from Arabidopsis thaliana
    Article Snippet: .. Plasmid DNA from transformed E. coli was linearized using the SacI restriction enzyme (Fermentas, Sankt Leon-Rot, Germany) overnight at 37°C. .. Approximately 10 μg of the linearized DNA was transformed into Pichia strain KM-71H cells by electroporation (Bio-Rad, Hercules, CA, USA) with following pulse settings: 25 μF, 1.5 kV, 200 Ω .

    Article Title: Entericidin Is Required for a Probiotic Treatment ( Enterobacter sp. Strain C6-6) To Protect Trout from Cold-Water Disease Challenge
    Article Snippet: .. The purified PCR template and the suicide vector pDM4 were both digested using the restriction enzymes SacI and XhoI for 3 h at 37°C, purified again, quantified by using a NanoDrop ND-1000 spectrophotometer (Thermo Scientific), and ligated using T4 DNA ligase (New England BioLabs). .. The ligated product was then transformed into electrocompetent E. coli S17 λ pir ( ) and selected on LB plates with chloramphenicol.

    Article Title: Cd36, a class B scavenger receptor, functions as a monomer to bind acetylated and oxidized low-density lipoproteins
    Article Snippet: .. The modified cDNA was excised from the plasmid using restriction enzymes SacI and AgeI and subcloned into similarly digested pIZ/V5-His (Invitrogen). .. This fused, in frame, the coding sequences for the 6-histidine tag of the cDNA with the 6-histidine tag of the pIZ/V5-His vector, thus encoding a protein of full-length Cd36 with a carboxy-terminal tag of GHHHHHHTGHHHHHH.

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  • 95
    Thermo Fisher unique saci site
    Unique Saci Site, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/unique saci site/product/Thermo Fisher
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    unique saci site - by Bioz Stars, 2020-09
    95/100 stars
      Buy from Supplier

    88
    Thermo Fisher saci xhoi sites
    Saci Xhoi Sites, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/saci xhoi sites/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    saci xhoi sites - by Bioz Stars, 2020-09
    88/100 stars
      Buy from Supplier

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