sfii restriction enzyme  (New England Biolabs)


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    Structured Review

    New England Biolabs sfii restriction enzyme
    Sfii Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sfii restriction enzyme/product/New England Biolabs
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    sfii restriction enzyme - by Bioz Stars, 2020-04
    90/100 stars

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    Clone Assay:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: Likewise, we sequentially fused LTI6b and T35S using the cloning sites EcoRI /SpeI , and SpeI /SfiI -B, respectively, to generate the C1 cassette. .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. Transformed cells were grown in super optimal broth supplemented with glucose and isolated clones were plated and the sequence confirmed by plasmid DNA sequencing.

    Amplification:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: The C1.1, C1.2, and C1.3 cassettes were generated by PCR amplification of the C1 cassette using pairs of oligonucleotide primers that replace the SfiI -A and SfiI -B with SfiI -C and SfiI -D, SfiI -E and SfiI -F, and SfiI -G and SfiI -H, respectively as indicated in Figure . .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    Lambda DNA Preparation:

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. For block 1, the voltage was 183 V, the initial switch time was 8 s, the final switch time was 20 s, and the duration was 20 h; for block 2, the voltage was 183 V, the initial switch time was 5 s, the final switch time was 8 s, and the duration was 20 h. The PFGE patterns were normalized with lambda DNA and low-range PFGE molecular weight markers (Bio-Rad).

    Blocking Assay:

    Article Title: Genotypic Characterization of Vibrio vulnificus Clinical Isolates in Korea
    Article Snippet: Lysed plugs were digested with NotI and SfiI (New England Biolabs, Boston, MA, USA), and PFGE was performed in 1% agarose gels in 0.5 × Tris–borate–EDTA buffer at 14°C using a CHEF mapper apparatus (Bio-Rad, Richmond, CA, USA) at 6 V/cm. .. Linearly ramped switching times were 3.5–50 seconds for 22 hours (used NotI) or block 1 second, 2–10 seconds for 13 hours and block 2, 20–25 seconds for 6 hours (used SfiI).

    Article Title: Coincident Resection at Both Ends of Random, ?-Induced Double-Strand Breaks Requires MRX (MRN), Sae2 (Ctp1), and Mre11-Nuclease
    Article Snippet: This section was cut from the full-length lane slices and processed for the 2nd dimension PFGE as follows: each lane section was equilibrated in 3 changes of 25 ml sterile TE (10 mM Tris, pH 8, 1 mM EDTA), treated with SfiI (New England Biolabs, Ipswich, MA; 400 units in 4 ml total reaction volume including gel slice) for 5 hr at 50°C, followed by overnight treatment in 1% sarkosyl (Sigma, St. Louis, MO) plus 1 mg/ml proteinase K (Invitrogen) at 37°C and subsequent equilibration with running buffer. .. After allowing the agarose to cool for 5 to 10 minutes, the end block was carefully removed, leaving the lane slices attached to the gel tray.

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. For block 1, the voltage was 183 V, the initial switch time was 8 s, the final switch time was 20 s, and the duration was 20 h; for block 2, the voltage was 183 V, the initial switch time was 5 s, the final switch time was 8 s, and the duration was 20 h. The PFGE patterns were normalized with lambda DNA and low-range PFGE molecular weight markers (Bio-Rad).

    Electrophoresis:

    Article Title: Genetic relationships between Candida albicans strains isolated from dental plaque, trachea, and bronchoalveolar lavage fluid from mechanically ventilated intensive care unit patients
    Article Snippet: For Sfi I digestion, the plug was transferred to a microcentrifuge tube containing 500 µL of buffer 2 (50 mM NaCl, 10 mMTris-HCl, 10 mM MgCl2 , 1 mM dithiothreitol), and incubated on ice for 30 min. After aspirating the buffer, the plug was placed in 160 µL (4X plug volume) of buffer 2 containing 20 U Sfi I and 2 µL BSA (New England BioLabs, Ipswich, MA), and incubated overnight at 50°C. .. The plugs were then placed into the agarose gel wells for electrophoresis.

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: .. The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. For block 1, the voltage was 183 V, the initial switch time was 8 s, the final switch time was 20 s, and the duration was 20 h; for block 2, the voltage was 183 V, the initial switch time was 5 s, the final switch time was 8 s, and the duration was 20 h. The PFGE patterns were normalized with lambda DNA and low-range PFGE molecular weight markers (Bio-Rad).

    Incubation:

    Article Title: Genetic relationships between Candida albicans strains isolated from dental plaque, trachea, and bronchoalveolar lavage fluid from mechanically ventilated intensive care unit patients
    Article Snippet: .. For Sfi I digestion, the plug was transferred to a microcentrifuge tube containing 500 µL of buffer 2 (50 mM NaCl, 10 mMTris-HCl, 10 mM MgCl2 , 1 mM dithiothreitol), and incubated on ice for 30 min. After aspirating the buffer, the plug was placed in 160 µL (4X plug volume) of buffer 2 containing 20 U Sfi I and 2 µL BSA (New England BioLabs, Ipswich, MA), and incubated overnight at 50°C. .. For Bss HII digestion, the plug was transferred to a microcentrifuge tube containing 500 µl of buffer 3 (100 mM NaCl, 50 mM Tris-HCl, 10 mM MgCl2 , 1 mM dithiothreitol), and incubated on ice for 30 min. After aspirating buffer, the plug was placed in 160 µl of buffer 3 containing 4 U of Bss HII (New England BioLabs), and incubated overnight at 50°C.

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany). .. For the SfiI cleavage, 0.5–1 μg DNA, 2 μl CutSmart® buffer and 0.2–0.5 μl SfiI (20 unit/μl) were added in 20 μl total volume, and then incubated for 1–16 h at 50°C in a thermocycler (MyCycler™, Bio-Rad, USA).

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: .. The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. For block 1, the voltage was 183 V, the initial switch time was 8 s, the final switch time was 20 s, and the duration was 20 h; for block 2, the voltage was 183 V, the initial switch time was 5 s, the final switch time was 8 s, and the duration was 20 h. The PFGE patterns were normalized with lambda DNA and low-range PFGE molecular weight markers (Bio-Rad).

    Modification:

    Article Title: Genotypic Characterization of Vibrio vulnificus Clinical Isolates in Korea
    Article Snippet: 2.3 PFGE Genetic relatedness between isolates recovered from individuals with V. vulnificus sepsis was investigated using PFGE as described by Gautom with some modification . .. Lysed plugs were digested with NotI and SfiI (New England Biolabs, Boston, MA, USA), and PFGE was performed in 1% agarose gels in 0.5 × Tris–borate–EDTA buffer at 14°C using a CHEF mapper apparatus (Bio-Rad, Richmond, CA, USA) at 6 V/cm.

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: The basic gene cassette (C1) modules were cloned in a pUC19 cloning vector (Clontech, Saint-Germain-en-Laye, France), which was modified to contain the SfiI -A and RsrII, SfiI -B recognition sites. .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. This modified PFGE protocol, which includes a significantly shorter method for making plugs (1 day instead of the usual 4 days), allowed us to complete the entire procedure (making the plugs and performing the electrophoresis) in approximately 4 days instead of the usual 7 days ( ).

    Transformation Assay:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. Transformed cells were grown in super optimal broth supplemented with glucose and isolated clones were plated and the sequence confirmed by plasmid DNA sequencing.

    Electroporation:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. The ligation product (pBAD33-eCPX-abltide) was desalted and electroporated into electrocompetent E. coli MC1061 with 2 mm electroporation cuvette and pulse at 2.5 kV, 50 µF, and 100 Ω.

    Southern Blot:

    Article Title: Cis-acting DNA sequence at a replication origin promotes repeat expansion to fragile X full mutation
    Article Snippet: The genomic DNA in the gel plugs was digested with 80 U SfiI (New England BioLabs, Inc.) at 50°C overnight. .. A Southern blot was performed to determine the location of the DNA fragment on the gel.

    Article Title: Factor VIII delivered by hematopoietic stem cell-derived B cells corrects the phenotype of hemophilia A mice
    Article Snippet: Southern blot analysis was performed as described previously ( , ). .. Genomic DNA (10 μg) was digested with Eco RI and Sfi I (New England Biolabs, Beverly, MA, USA), separated through 0.8% agarose gels, and transferred to Hybond-N+ membranes (Amersham Biosciences, Piscataway, NJ, USA).

    Ligation:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: Meanwhile, four binary vector backbones were PCR-amplified from the plasmid pXNS2pat-YFP (accession number KF499077; Dahncke and Witte, ) using four pairs of primers that add distinct SfiI cleavage sites in the flanks of the backbones to specifically allow ligation with the overhangs of either C1, C1.1, C1.2, or C1.3 gene cassette as indicated in Supplementary Figure . .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. The ligation product (pBAD33-eCPX-abltide) was desalted and electroporated into electrocompetent E. coli MC1061 with 2 mm electroporation cuvette and pulse at 2.5 kV, 50 µF, and 100 Ω.

    Generated:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: The C1.1, C1.2, and C1.3 cassettes were generated by PCR amplification of the C1 cassette using pairs of oligonucleotide primers that replace the SfiI -A and SfiI -B with SfiI -C and SfiI -D, SfiI -E and SfiI -F, and SfiI -G and SfiI -H, respectively as indicated in Figure . .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    other:

    Article Title: Evolution and Spread of a Multidrug-Resistant Proteus mirabilis Clone with Chromosomal AmpC-Type Cephalosporinases in Europe ▿
    Article Snippet: For PFGE, total P. mirabilis DNA in agarose plugs was prepared as described previously , digested with NotI and SfiI (New England Biolabs), and electrophoresed using a CHEF III Bio-Rad apparatus (Hercules, CA).

    DNA Sequencing:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. Transformed cells were grown in super optimal broth supplemented with glucose and isolated clones were plated and the sequence confirmed by plasmid DNA sequencing.

    Polymerase Chain Reaction:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: Meanwhile, four binary vector backbones were PCR-amplified from the plasmid pXNS2pat-YFP (accession number KF499077; Dahncke and Witte, ) using four pairs of primers that add distinct SfiI cleavage sites in the flanks of the backbones to specifically allow ligation with the overhangs of either C1, C1.1, C1.2, or C1.3 gene cassette as indicated in Supplementary Figure . .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany).

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: .. The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. The ligation product (pBAD33-eCPX-abltide) was desalted and electroporated into electrocompetent E. coli MC1061 with 2 mm electroporation cuvette and pulse at 2.5 kV, 50 µF, and 100 Ω.

    Pulsed-Field Gel:

    Article Title: Cis-acting DNA sequence at a replication origin promotes repeat expansion to fragile X full mutation
    Article Snippet: The genomic DNA in the gel plugs was digested with 80 U SfiI (New England BioLabs, Inc.) at 50°C overnight. .. A gel λ ladder pulsed-field gel electrophoresis (PFGE) marker and yeast chromosome PFGE marker (both obtained from New England BioLabs, Inc.) were cast next to the gel plugs.

    Migration:

    Article Title: Genotypic Characterization of Vibrio vulnificus Clinical Isolates in Korea
    Article Snippet: Lysed plugs were digested with NotI and SfiI (New England Biolabs, Boston, MA, USA), and PFGE was performed in 1% agarose gels in 0.5 × Tris–borate–EDTA buffer at 14°C using a CHEF mapper apparatus (Bio-Rad, Richmond, CA, USA) at 6 V/cm. .. Analysis of banding patterns was performed using the Dice coefficient with a 1.0% tolerance for the band migration distance.

    Article Title: Coincident Resection at Both Ends of Random, ?-Induced Double-Strand Breaks Requires MRX (MRN), Sae2 (Ctp1), and Mre11-Nuclease
    Article Snippet: This procedure avoided the need to stain the lanes prior to the 2nd run, since the stain interfered with the migration of chromosome fragments in the 2nd dimension. .. This section was cut from the full-length lane slices and processed for the 2nd dimension PFGE as follows: each lane section was equilibrated in 3 changes of 25 ml sterile TE (10 mM Tris, pH 8, 1 mM EDTA), treated with SfiI (New England Biolabs, Ipswich, MA; 400 units in 4 ml total reaction volume including gel slice) for 5 hr at 50°C, followed by overnight treatment in 1% sarkosyl (Sigma, St. Louis, MO) plus 1 mg/ml proteinase K (Invitrogen) at 37°C and subsequent equilibration with running buffer.

    Radioactivity:

    Article Title: Factor VIII delivered by hematopoietic stem cell-derived B cells corrects the phenotype of hemophilia A mice
    Article Snippet: Genomic DNA (10 μg) was digested with Eco RI and Sfi I (New England Biolabs, Beverly, MA, USA), separated through 0.8% agarose gels, and transferred to Hybond-N+ membranes (Amersham Biosciences, Piscataway, NJ, USA). .. Digital images were acquired using a Storm 860 PhosphorImager and radioactivity was quantitated using ImageQuant software (Amersham Biosciences).

    Isolation:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. Transformed cells were grown in super optimal broth supplemented with glucose and isolated clones were plated and the sequence confirmed by plasmid DNA sequencing.

    Sequencing:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. Transformed cells were grown in super optimal broth supplemented with glucose and isolated clones were plated and the sequence confirmed by plasmid DNA sequencing.

    Construct:

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: Paragraph title: Bacterial surface display construct ... The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated.

    Lysis:

    Article Title: Herpesvirus telomeric repeats facilitate genomic integration into host telomeres and mobilization of viral DNA during reactivation
    Article Snippet: .. 1 × 107 /ml LCLs were embedded in 1% agarose and digested at 50°C for 48 h hours in lysis buffer (0.5 M EDTA and 1% wt/vol N -laurylsarcosine) containing 1 mg/ml proteinase K. Proteinase K was inactivated with 0.01 mM phenylmethanesulfonyl fluoride, and agarose plugs were digested with either SfiI or BclI (New England Biolabs) overnight according to the manufacturer’s instructions. .. DNA fragments were resolved via PFGE using the CHEF-DR III system (Bio-Rad Laboratories).

    Plasmid Preparation:

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany). .. For the SfiI cleavage, 0.5–1 μg DNA, 2 μl CutSmart® buffer and 0.2–0.5 μl SfiI (20 unit/μl) were added in 20 μl total volume, and then incubated for 1–16 h at 50°C in a thermocycler (MyCycler™, Bio-Rad, USA).

    Article Title: A Novel Quantitative Kinase Assay Using Bacterial Surface Display and Flow Cytometry
    Article Snippet: .. The product of PCR reaction and the vector pBAD33-eCPX were digested with SFiI (New England Biolabs) and ligated. .. The ligation product (pBAD33-eCPX-abltide) was desalted and electroporated into electrocompetent E. coli MC1061 with 2 mm electroporation cuvette and pulse at 2.5 kV, 50 µF, and 100 Ω.

    Software:

    Article Title: Nosocomial Pneumonia Caused by Three Genetically Different Strains of Legionella pneumophila and Detection of These Strains in the Hospital Water Supply
    Article Snippet: For macrorestriction analysis (MRA), chromosomal DNAs were digested overnight with Sfi I, Asc I, and Not I (New England Biolabs, Schwahlbach, Germany) and separated with the CHEF III System (BioRad Laboratories, Munich, Germany) ( ). .. Computer-assisted analysis of the restriction patterns was performed with the software package GelCompare (Applied Maths, Kortrijk, Belgium).

    Article Title: Genotypic Characterization of Vibrio vulnificus Clinical Isolates in Korea
    Article Snippet: Lysed plugs were digested with NotI and SfiI (New England Biolabs, Boston, MA, USA), and PFGE was performed in 1% agarose gels in 0.5 × Tris–borate–EDTA buffer at 14°C using a CHEF mapper apparatus (Bio-Rad, Richmond, CA, USA) at 6 V/cm. .. PFGE banding pattern analysis was performed using BioNumerics software (version 4.6; Applied Math, Kortrijk, Belgium).

    Article Title: Factor VIII delivered by hematopoietic stem cell-derived B cells corrects the phenotype of hemophilia A mice
    Article Snippet: Genomic DNA (10 μg) was digested with Eco RI and Sfi I (New England Biolabs, Beverly, MA, USA), separated through 0.8% agarose gels, and transferred to Hybond-N+ membranes (Amersham Biosciences, Piscataway, NJ, USA). .. Digital images were acquired using a Storm 860 PhosphorImager and radioactivity was quantitated using ImageQuant software (Amersham Biosciences).

    Agarose Gel Electrophoresis:

    Article Title: Cis-acting DNA sequence at a replication origin promotes repeat expansion to fragile X full mutation
    Article Snippet: The genomic DNA in the gel plugs was digested with 80 U SfiI (New England BioLabs, Inc.) at 50°C overnight. .. The digested gel plugs were rinsed with TE and cast into a 0.7% SeaPlaque GTG agarose gel (Lonza).

    Article Title: Genetic relationships between Candida albicans strains isolated from dental plaque, trachea, and bronchoalveolar lavage fluid from mechanically ventilated intensive care unit patients
    Article Snippet: For Sfi I digestion, the plug was transferred to a microcentrifuge tube containing 500 µL of buffer 2 (50 mM NaCl, 10 mMTris-HCl, 10 mM MgCl2 , 1 mM dithiothreitol), and incubated on ice for 30 min. After aspirating the buffer, the plug was placed in 160 µL (4X plug volume) of buffer 2 containing 20 U Sfi I and 2 µL BSA (New England BioLabs, Ipswich, MA), and incubated overnight at 50°C. .. The plugs were then placed into the agarose gel wells for electrophoresis.

    Article Title: COLORFUL-Circuit: A Platform for Rapid Multigene Assembly, Delivery, and Expression in Plants
    Article Snippet: .. The gene cassettes C1, C1.1, C1.2, or C1.3 and their corresponding vector backbones were cleaved with SfiI (NEB, Germany), separated on agarose gel electrophoresis, and then cleaned up from the gel using innuPREP DOUBLEpure Kit (Analytik Jena, Germany). .. For the SfiI cleavage, 0.5–1 μg DNA, 2 μl CutSmart® buffer and 0.2–0.5 μl SfiI (20 unit/μl) were added in 20 μl total volume, and then incubated for 1–16 h at 50°C in a thermocycler (MyCycler™, Bio-Rad, USA).

    In Vitro:

    Article Title: Experimental surgery to create subgenomes of Bacillus subtilis 168
    Article Snippet: Paragraph title: In Vitro DNA Manipulations. ... Endonucleases and T4 DNA ligase were obtained from Toyobo (Osaka), except for Sfi I and I- Ceu I from New England Biolabs, Not I from Takara Shuzo (Kyoto), and I- Sce I from Boehringer Mannheim.

    Article Title: Coincident Resection at Both Ends of Random, ?-Induced Double-Strand Breaks Requires MRX (MRN), Sae2 (Ctp1), and Mre11-Nuclease
    Article Snippet: This section was cut from the full-length lane slices and processed for the 2nd dimension PFGE as follows: each lane section was equilibrated in 3 changes of 25 ml sterile TE (10 mM Tris, pH 8, 1 mM EDTA), treated with SfiI (New England Biolabs, Ipswich, MA; 400 units in 4 ml total reaction volume including gel slice) for 5 hr at 50°C, followed by overnight treatment in 1% sarkosyl (Sigma, St. Louis, MO) plus 1 mg/ml proteinase K (Invitrogen) at 37°C and subsequent equilibration with running buffer. .. The map of the positions of the SfiI restriction sites relative to the I-Sce I cut site are presented in and results for cutting the DNA by I-Sce I in vitro (prior to 1st dimension) and SfiI (prior to 2nd dimension) are presented in .

    Concentration Assay:

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: The cell suspension was diluted to an optical density at 610 nm of 14 to 15, and proteinase K (2 mg/ml, final concentration) was added to the suspension. .. The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer.

    Molecular Weight:

    Article Title: Diphtheria in the Republic of Georgia: Use of Molecular Typing Techniques for Characterization of Corynebacterium diphtheriae Strains
    Article Snippet: The plugs were incubated (30 min, room temperature) with restriction enzyme buffer, the DNA in the plugs was digested by incubation (50°C, 5 h) of the plugs with Sfi I (New England Biolabs, Beverly, Mass.), and electrophoresis was performed with 1% SeaKem Gold agarose in 0.5× Tris-borate-EDTA buffer. .. For block 1, the voltage was 183 V, the initial switch time was 8 s, the final switch time was 20 s, and the duration was 20 h; for block 2, the voltage was 183 V, the initial switch time was 5 s, the final switch time was 8 s, and the duration was 20 h. The PFGE patterns were normalized with lambda DNA and low-range PFGE molecular weight markers (Bio-Rad).

    Marker:

    Article Title: Cis-acting DNA sequence at a replication origin promotes repeat expansion to fragile X full mutation
    Article Snippet: The genomic DNA in the gel plugs was digested with 80 U SfiI (New England BioLabs, Inc.) at 50°C overnight. .. A gel λ ladder pulsed-field gel electrophoresis (PFGE) marker and yeast chromosome PFGE marker (both obtained from New England BioLabs, Inc.) were cast next to the gel plugs.

    Staining:

    Article Title: Coincident Resection at Both Ends of Random, ?-Induced Double-Strand Breaks Requires MRX (MRN), Sae2 (Ctp1), and Mre11-Nuclease
    Article Snippet: After staining the flanks for 10 minutes in SYBR Gold, they were digitally photographed, and a ruler was used in the photograph to identify the region of the unstained lane slice corresponding to ∼220 kb to 600 kb (from just below Chr I to just above Chr V). .. This section was cut from the full-length lane slices and processed for the 2nd dimension PFGE as follows: each lane section was equilibrated in 3 changes of 25 ml sterile TE (10 mM Tris, pH 8, 1 mM EDTA), treated with SfiI (New England Biolabs, Ipswich, MA; 400 units in 4 ml total reaction volume including gel slice) for 5 hr at 50°C, followed by overnight treatment in 1% sarkosyl (Sigma, St. Louis, MO) plus 1 mg/ml proteinase K (Invitrogen) at 37°C and subsequent equilibration with running buffer.

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    New England Biolabs sfii restriction enzyme
    Sfii Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sfii restriction enzyme/product/New England Biolabs
    Average 90 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    sfii restriction enzyme - by Bioz Stars, 2020-04
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