sbfi  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    SbfI
    Description:
    SbfI 2 500 units
    Catalog Number:
    R0642L
    Price:
    297
    Size:
    2 500 units
    Category:
    Restriction Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs sbfi
    SbfI
    SbfI 2 500 units
    https://www.bioz.com/result/sbfi/product/New England Biolabs
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    sbfi - by Bioz Stars, 2019-07
    99/100 stars

    Images

    Related Articles

    Transduction:

    Article Title: Early Passage Mesenchymal Stem Cells Display Decreased Radiosensitivity and Increased DNA Repair Activity
    Article Snippet: For overexpression of PARP‐1, MSCs were transduced with PLAS2w vector carrying full length PARP‐1 gene from National Yang‐Ming University VYM Genome Research Center. .. In brief, PLAS2w plasmid was digested by SbfI/AgeI restriction enzymes (New England Biolabs; Ipswich MA; https://www.neb.com /) and the full length of PARP‐1 gene was ligased by T7 DNA ligase (New England Biolabs).

    Article Title: Impacts of Global Transcriptional Regulators on Persister Metabolism
    Article Snippet: Standard P1 phage transduction was employed to transfer sequences with the genetic deletions (Δ arcA , Δ cra , Δ crp , Δ dksA , Δ fnr , Δ lrp , and Δ rpoS ) from the Keio Collection to E. coli MG1655 ( ). .. The amplified genes were digested with XhoI and SbfI (New England BioLabs, Ipswich, MA) and cloned into pUA66 ( ).

    Clone Assay:

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: Next, the full-length fragment, including the promoter and terminator, was PCR amplified from p94FDH and cloned into the NotI (NEB)-linearized and blunt-ended pKOSIGK_mazF plasmid, yielding plasmid pKISIGK::FDH_mazF. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL.

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs). .. The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs).

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The human ERα coding gene in pSNAP-hERα vector was lifted from pJ3-FLAG-hERα (a gift from Dr. Carolyn L. Smith, Baylor College of Medicine). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ). .. Reporter vectors 3×ERE TATA luc (Addgene plasmid 11354), 2×PRE TK luc (Addgene plasmid 11350), and pGL3 luc (Promega) all contained a firefly ( Photinus pyralis ) luciferase gene.

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Recoded HIV-1 proteases and wild-type HXB2 protease were cloned into the GFP reporter HIV-1 infectious clone pNL4-3-deltaE-EGFP (NIH AIDS Research and Reference Reagent Program). .. Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Recombinant pNL4-3-deltaE-EGFP plasmids (4 μg) were used to transfect 8 × 105 293 T cells in the presence of Lipofectamine 2000 (Invitrogen).

    Article Title: Impacts of Global Transcriptional Regulators on Persister Metabolism
    Article Snippet: To complement Δ crp and Δ cyaA , the native promoters and genes were amplified from E. coli MG1655 genomic DNA using primers 5′-CTAGTAGCTCGAGTTTTGCTACTCCACTGCGTC-3′ and 5′-GCATCATCCTGCAGGTTAACGAGTGCCGTAAACGA-3′ for crp and primers 5′-CTAGTAGCTCGAGAGTGTGCCTGCCAGAGTGCA-3′ and 5′-GCATCATCCTGCAGGTCACGAAAAATATTGCTGTA-3′ for cyaA . .. The amplified genes were digested with XhoI and SbfI (New England BioLabs, Ipswich, MA) and cloned into pUA66 ( ). .. All gene deletion and cloning constructions were confirmed by colony PCR and/or DNA sequencing (Genewiz, South Plainfield, NJ).

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ).

    Amplification:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: The RAD specific P1 and P2 paired-end adapters and library amplification PCR primer sequences used in this study are detailed in Baxter et al. . .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: BAsE-Seq: a method for obtaining long viral haplotypes from short sequence reads
    Article Snippet: Excess primers were removed by Exonuclease I (Enzymatics, Beverly, MA, USA). .. We clonally amplified 40,000 copies of barcode-tagged genomes using the Long PCR Enzyme Mix (Thermo Scientific) and universal primers, then digested with SbfI (NEB) to protect the barcode-proximal end from exonuclease digest. .. Next, unidirectional nested deletions from the barcode-distal end were generated using a combination of Exonuclease III and S1 Nuclease (Promega) to achieve a broad size distribution of fragments ranging from approximately 300 bp to 3,200 bp.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: The RAD-specific P1 and P2 paired-end adapters and library amplification PCR primer sequences used in this study are detailed in . .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: Next, the full-length fragment, including the promoter and terminator, was PCR amplified from p94FDH and cloned into the NotI (NEB)-linearized and blunt-ended pKOSIGK_mazF plasmid, yielding plasmid pKISIGK::FDH_mazF. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL.

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Recoded HIV-1 proteases and wild-type HXB2 protease were cloned into the GFP reporter HIV-1 infectious clone pNL4-3-deltaE-EGFP (NIH AIDS Research and Reference Reagent Program). .. Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Recombinant pNL4-3-deltaE-EGFP plasmids (4 μg) were used to transfect 8 × 105 293 T cells in the presence of Lipofectamine 2000 (Invitrogen).

    Article Title: Amplification Biases and Consistent Recovery of Loci in a Double-Digest RAD-seq Protocol
    Article Snippet: We then double-digest ∼1.0 µg of DNA with high fidelity versions of the SbfI and EcoRI restriction enzymes (New England Biolabs); when less DNA is available, we have had good success starting with as little as 0.17 µg of genomic DNA. .. Following digestion, ligation of double-stranded sequencing adapters is completed in the same tube.

    Article Title: Impacts of Global Transcriptional Regulators on Persister Metabolism
    Article Snippet: To complement Δ crp and Δ cyaA , the native promoters and genes were amplified from E. coli MG1655 genomic DNA using primers 5′-CTAGTAGCTCGAGTTTTGCTACTCCACTGCGTC-3′ and 5′-GCATCATCCTGCAGGTTAACGAGTGCCGTAAACGA-3′ for crp and primers 5′-CTAGTAGCTCGAGAGTGTGCCTGCCAGAGTGCA-3′ and 5′-GCATCATCCTGCAGGTCACGAAAAATATTGCTGTA-3′ for cyaA . .. The amplified genes were digested with XhoI and SbfI (New England BioLabs, Ipswich, MA) and cloned into pUA66 ( ). .. All gene deletion and cloning constructions were confirmed by colony PCR and/or DNA sequencing (Genewiz, South Plainfield, NJ).

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ).

    Expressing:

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: All constructs were in the pGEMhe vector, a cDNA plasmid optimized for protein expression in Xenopus oocytes. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The His6 -tagged ERα DNA binding domain (ERα-DBD) expression construct was a gift from Dr. W. Lee Kraus (University of Texas Southwestern Medical Center). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ).

    Cytometry:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Replication capacity is expressed as the percentage of p24 antigen produced by the vectors containing each of the derived protease sequences compared to the p24 antigen from the vector containing the HIV-1 HXB2 protease reference sequence (100%).

    Construct:

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: RAD libraries were constructed following a protocol from Baird et al. ( ). .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: Next, the full-length fragment, including the promoter and terminator, was PCR amplified from p94FDH and cloned into the NotI (NEB)-linearized and blunt-ended pKOSIGK_mazF plasmid, yielding plasmid pKISIGK::FDH_mazF. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL. .. The lactose-inducible promoter and its divergent regulator ( bgaR -PbgaL ) were utilized and fused immediately upstream of a 100-bp antisense RNA (asRNA) that targets the first 100 bp of the repL transcript coding for the replication protein (RepL), creating plasmid p94FLP_asRepL.

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: All constructs were in the pGEMhe vector, a cDNA plasmid optimized for protein expression in Xenopus oocytes. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: We constructed 14 novel RADseq libraries, each consisting of between 24 and 36 pooled, individually labeled O. nerka individuals (n = 408 total) following as modified in ). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The His6 -tagged ERα DNA binding domain (ERα-DBD) expression construct was a gift from Dr. W. Lee Kraus (University of Texas Southwestern Medical Center). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ).

    Article Title: Phenotype of the Most Common "Slow Acetylator" Arylamine N-Acetyltransferase 1 Genetic Variant (NAT1*14B) Is Substrate-Dependent
    Article Snippet: The NATb /NAT1 * 4 construct was then ligated into pcDNA5/FRT using T4 ligase (New England Biolabs). .. To construct the NATb/ NAT1 * 14B pcDNA5/FRT plasmid, the NATb/ NAT1 * 4 pcDNA5/FRT and a previously constructed NAT1 * 14B allelic construct expressed in a yeast vector, pESP-3 (Agilent Technologies) , were both incubated at 37°C with restriction enzymes SbfI and AflII (New England Biolabs). .. After restriction digestion, the NATb/ NAT1 * 4 pcDNA5/FRT and the 476-base pair segment of NAT1 * 14B (including G560A) were gel purified and ligated using T4 ligase (New England Biolabs).

    Electrophoresis:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop) and quality assessed by agarose gel electrophoresis, and was finally diluted to a concentration of 50 ng/µL in 5 mmol/L Tris, pH 8.5. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Incubation:

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C. .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Phenotype of the Most Common "Slow Acetylator" Arylamine N-Acetyltransferase 1 Genetic Variant (NAT1*14B) Is Substrate-Dependent
    Article Snippet: The NATb /NAT1 * 4 construct was then ligated into pcDNA5/FRT using T4 ligase (New England Biolabs). .. To construct the NATb/ NAT1 * 14B pcDNA5/FRT plasmid, the NATb/ NAT1 * 4 pcDNA5/FRT and a previously constructed NAT1 * 14B allelic construct expressed in a yeast vector, pESP-3 (Agilent Technologies) , were both incubated at 37°C with restriction enzymes SbfI and AflII (New England Biolabs). .. After restriction digestion, the NATb/ NAT1 * 4 pcDNA5/FRT and the 476-base pair segment of NAT1 * 14B (including G560A) were gel purified and ligated using T4 ligase (New England Biolabs).

    Luciferase:

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ). .. Vector 3×ERE TATA luc had three copies of Xenopus laevis vitellogenin A2 ERE in the reporter gene promoter (Hall and McDonnell, ); 2×PRE TK luc contained two copies of a consensus progesterone response element (PRE) upstream of the thymidine kinase promoter (Giangrande et al., ).

    Activity Assay:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Replication capacity is expressed as the percentage of p24 antigen produced by the vectors containing each of the derived protease sequences compared to the p24 antigen from the vector containing the HIV-1 HXB2 protease reference sequence (100%).

    Infection:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Paragraph title: Single-cycle infectivity assay ... Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites.

    Mass Spectrometry:

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion). .. The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion).

    Modification:

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: Total genomic DNA was extracted from each sample using the modified CTAB method as described by Doyle and Doyle ( ). .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: We constructed 14 novel RADseq libraries, each consisting of between 24 and 36 pooled, individually labeled O. nerka individuals (n = 408 total) following as modified in ). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Transformation Assay:

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs). .. The 7.3kb fragment was purified from a 1% agarose gel using the QIAquick Gel Extraction Kit (Qiagen).

    Over Expression:

    Article Title: Early Passage Mesenchymal Stem Cells Display Decreased Radiosensitivity and Increased DNA Repair Activity
    Article Snippet: Paragraph title: Overexpression of PARP‐1 in MSCs ... In brief, PLAS2w plasmid was digested by SbfI/AgeI restriction enzymes (New England Biolabs; Ipswich MA; https://www.neb.com /) and the full length of PARP‐1 gene was ligased by T7 DNA ligase (New England Biolabs).

    Derivative Assay:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. The relative replication capacity of the virus was determined by measuring the amount of p24 antigen produced 72 h after transfection.

    Article Title: Impacts of Global Transcriptional Regulators on Persister Metabolism
    Article Snippet: All strains were derived from Escherichia coli MG1655. .. The amplified genes were digested with XhoI and SbfI (New England BioLabs, Ipswich, MA) and cloned into pUA66 ( ).

    Gel Purification:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB). .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Transfection:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites.

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ).

    Gas Chromatography:

    Article Title: Amplification Biases and Consistent Recovery of Loci in a Double-Digest RAD-seq Protocol
    Article Snippet: We then double-digest ∼1.0 µg of DNA with high fidelity versions of the SbfI and EcoRI restriction enzymes (New England Biolabs); when less DNA is available, we have had good success starting with as little as 0.17 µg of genomic DNA. .. The P1 adapter includes the Illumina TruSeq forward amplification and sequencing primer sequences, one of 48 unique, six bp barcodes, and a TGCA overhang on the top strand to match the sticky end left by SbfI (Table S2 in ).

    Ligation:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: BAsE-Seq: a method for obtaining long viral haplotypes from short sequence reads
    Article Snippet: We clonally amplified 40,000 copies of barcode-tagged genomes using the Long PCR Enzyme Mix (Thermo Scientific) and universal primers, then digested with SbfI (NEB) to protect the barcode-proximal end from exonuclease digest. .. Barcode-containing fragments were purified using streptavidin-coated Dynabeads (Life Technologies) and subjected to end repair using T4 DNA polymerase and T4 Polynucleotide Kinase (NEB).

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C. .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB). .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Introduce:

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Generated:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Replication capacity is expressed as the percentage of p24 antigen produced by the vectors containing each of the derived protease sequences compared to the p24 antigen from the vector containing the HIV-1 HXB2 protease reference sequence (100%).

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: Point mutations of hERG were generated by PCR using the overlap extension technique ( ). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ).

    other:

    Article Title: An efficient method of directly cloning chimpanzee adenovirus as a vaccine vector
    Article Snippet: EcoRI (New England Biolabs, cat. no. R0101S) KpnI (New England Biolabs, cat. no. R0142S) PmeI (New England Biolabs, cat. no. R0560S) PacI (New England Biolabs, cat. no. R0547S) XbaI (New England Biolabs, cat. no. R0145S) SnaBI (New England Biolabs, cat. no. R0130S) NdeI (New England Biolabs, cat. no. R0111S) SphI (New England Biolabs, cat. no. R0182S) HindIII (New England Biolabs, cat. no. R0104S) AgeI (New England Biolabs, cat. no. R0552S) MluI (New England Biolabs, cat. no. R0198S) SbfI (New England Biolabs, cat. no. R0642S) BstAPI (New England Biolabs, cat. no. V0269S) SwaI (New England Biolabs, cat. no. R0604S) I-CeuI (New England Biolabs, cat. no. R0699S) PI-SceI (New England Biolabs, cat. no. R0696S) SgfI (Promega, cat. no. R5104) Eco47III (Promega, cat. no. R6731) Phusion hot start high-fidelity DNA polymerase (England Biolabs, cat. no. F-540S) dNTP mix (10 mM each; Invitrogen, cat. no. R72501) Alkaline phosphatase (AP; Roche, cat. no. 13826120) T4 DNA ligase (Roche, cat. no. 13827621) DNA ladders (Invitrogen, cat. nos.

    Overlap Extension Polymerase Chain Reaction:

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL.

    DNA Sequencing:

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: Restriction-site associated DNA (RAD) strategy combined with Illumina DNA sequencing was used for the fast and effective identification of SNP markers. .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Polymerase Chain Reaction:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: The RAD specific P1 and P2 paired-end adapters and library amplification PCR primer sequences used in this study are detailed in Baxter et al. . .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: BAsE-Seq: a method for obtaining long viral haplotypes from short sequence reads
    Article Snippet: Excess primers were removed by Exonuclease I (Enzymatics, Beverly, MA, USA). .. We clonally amplified 40,000 copies of barcode-tagged genomes using the Long PCR Enzyme Mix (Thermo Scientific) and universal primers, then digested with SbfI (NEB) to protect the barcode-proximal end from exonuclease digest. .. Next, unidirectional nested deletions from the barcode-distal end were generated using a combination of Exonuclease III and S1 Nuclease (Promega) to achieve a broad size distribution of fragments ranging from approximately 300 bp to 3,200 bp.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: The RAD-specific P1 and P2 paired-end adapters and library amplification PCR primer sequences used in this study are detailed in . .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: Next, the full-length fragment, including the promoter and terminator, was PCR amplified from p94FDH and cloned into the NotI (NEB)-linearized and blunt-ended pKOSIGK_mazF plasmid, yielding plasmid pKISIGK::FDH_mazF. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL.

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: Site-directed mutagenesis was performed by PCR using the Stratagene QuikChange protocol, and primers ordered from Integrated DNA Technologies. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Recoded HIV-1 proteases and wild-type HXB2 protease were cloned into the GFP reporter HIV-1 infectious clone pNL4-3-deltaE-EGFP (NIH AIDS Research and Reference Reagent Program). .. Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Recombinant pNL4-3-deltaE-EGFP plasmids (4 μg) were used to transfect 8 × 105 293 T cells in the presence of Lipofectamine 2000 (Invitrogen).

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. A QuikChange II XL Site-Directed Mutagenesis Kit (Agilent Technologies, Santa Clara, CA, USA) was used to introduce an SbfI restriction site at position 6849 (downstream from the polyA term site), and an SgrDI restriction site at position 4320 (3′ end of the puromycin resistance cassette) of the pAAVS1-puro-DNR vector. pAAVS1-puro-DNR was digested with SbfI/SgrDI (ThermoFisher Scientific) and ligated with the 4434bp SbfI/XhoI pROSA26-1 fragment to form the pAAVS1-ROSA26 vector. pAAVS1-ROSA26 was digested with SgfI/MluI (NEB) to release a 7054bp product which was purified by electrophoresis as above.

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ).

    Sonication:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Injection:

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion). .. Acylation of tRNA was confirmed using MALDI mass spectrometry using a 3-hydroxypicolinic acid matrix.

    Binding Assay:

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: This α4L’Aβ2 construct is described as wild type and/or α4β2 throughout the report for clarity in comparing noncanonical mutations made to the binding site, which is over 60 Å away from the channel gate. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The His6 -tagged ERα DNA binding domain (ERα-DBD) expression construct was a gift from Dr. W. Lee Kraus (University of Texas Southwestern Medical Center). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ).

    DNA Extraction:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: DNA was extracted from fin tissue of the fish using the REALPure genomic DNA extraction kit (Durviz S.L.) and treated with RNase to remove residual RNA from the sample. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Genomic DNA was extracted from fin samples using the REALPure genomic DNA extraction kit (Durviz S.L.) and treated with RNase. .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Mutagenesis:

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL.

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: Mutagenesis of the GluClβ receptor was performed using the QuikChange protocol (Stratagene), and for nonsense suppression experiments, a TAG codon was mutated into the site of interest. .. The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion).

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: Paragraph title: CD13 enzymatic mutation ... The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs).

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: Site-directed mutagenesis was performed by PCR using the Stratagene QuikChange protocol, and primers ordered from Integrated DNA Technologies. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: The final PCR product was cloned into Zero Blunt Vector (Invitrogen). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. All mutations were verified using a high-throughput 48 capillary ABI 3730 sequencer (UCDNA Services, University of Calgary). hERG mutant channels were transiently expressed in HEK 293 cells (American Type Culture Collection).

    Isolation:

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs). .. The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs).

    Flow Cytometry:

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Replication capacity is expressed as the percentage of p24 antigen produced by the vectors containing each of the derived protease sequences compared to the p24 antigen from the vector containing the HIV-1 HXB2 protease reference sequence (100%).

    Labeling:

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: We constructed 14 novel RADseq libraries, each consisting of between 24 and 36 pooled, individually labeled O. nerka individuals (n = 408 total) following as modified in ). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Purification:

    Article Title: BAsE-Seq: a method for obtaining long viral haplotypes from short sequence reads
    Article Snippet: We clonally amplified 40,000 copies of barcode-tagged genomes using the Long PCR Enzyme Mix (Thermo Scientific) and universal primers, then digested with SbfI (NEB) to protect the barcode-proximal end from exonuclease digest. .. Next, unidirectional nested deletions from the barcode-distal end were generated using a combination of Exonuclease III and S1 Nuclease (Promega) to achieve a broad size distribution of fragments ranging from approximately 300 bp to 3,200 bp.

    Article Title: Efficient Production of Papillomavirus Gene Delivery Vectors in Defined In Vitro Reactions
    Article Snippet: All pVITRO constructs were a kind gift from Richard Roden (Department of Pathology, The Johns Hopkins University) and have been described previously by Kwak et al. .. All plasmids were produced in competent Escherichia coli DH5α (BIO-85026, Bioline) and purified using the Plasmid Plus Midi Kit (12945, QIAGEN). pfwB was linearized using SbfI (R3642, New England Biolabs) and pCLucf with XmnI (R0194, New England Biolabs). pCLucf was digested with EcoRI (R3101, New England Biolabs) for the linear split virus used in . .. Digestion was confirmed by agarose gel electrophoresis, and enzymes were heat-inactivated according to the manufacturer’s instructions before use.

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: Site-directed mutagenesis was performed by PCR using the Stratagene QuikChange protocol, and primers ordered from Integrated DNA Technologies. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN). .. Final concentrations were quantified by UV spectroscopy.

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The protein was expressed in E.coli BL21(DE3) strain, and purified using ProBond™ Resin (Invitrogen) according to the manufacturer's instructions. .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ).

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product. .. pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Sequencing:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Paragraph title: RAD Library Preparation and Sequencing ... Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: BAsE-Seq: a method for obtaining long viral haplotypes from short sequence reads
    Article Snippet: Using HBV-specific primers that contain universal sequence on their 5′ ends, 106 HBV genomes were uniquely assigned to a molecular barcode (20 random nucleotides) by performing two cycles of PCR using the Long PCR Enzyme Mix (Thermo Scientific, Waltham, MA, USA). .. We clonally amplified 40,000 copies of barcode-tagged genomes using the Long PCR Enzyme Mix (Thermo Scientific) and universal primers, then digested with SbfI (NEB) to protect the barcode-proximal end from exonuclease digest.

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: Paragraph title: RAD library construction and sequencing ... The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Paragraph title: RAD Library Preparation and Sequencing ... Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: A CD13 clone (MGC Human ANPEP Sequence-Verified cDNA, ) was obtained from Open Biosystems (Thermo Scientific). .. The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs).

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The human ERα coding gene in pSNAP-hERα vector was lifted from pJ3-FLAG-hERα (a gift from Dr. Carolyn L. Smith, Baylor College of Medicine). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ). .. Reporter vectors 3×ERE TATA luc (Addgene plasmid 11354), 2×PRE TK luc (Addgene plasmid 11350), and pGL3 luc (Promega) all contained a firefly ( Photinus pyralis ) luciferase gene.

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. The relative replication capacity of the virus was determined by measuring the amount of p24 antigen produced 72 h after transfection.

    Article Title: Amplification Biases and Consistent Recovery of Loci in a Double-Digest RAD-seq Protocol
    Article Snippet: We then double-digest ∼1.0 µg of DNA with high fidelity versions of the SbfI and EcoRI restriction enzymes (New England Biolabs); when less DNA is available, we have had good success starting with as little as 0.17 µg of genomic DNA. .. Following digestion, ligation of double-stranded sequencing adapters is completed in the same tube.

    Article Title: Processes Driving the Adaptive Radiation of a Tropical Tree (Diospyros, Ebenaceae) in New Caledonia, a Biodiversity Hotspot
    Article Snippet: By using an average genome size in the target group of 1C = 1.9 pg ( ) and the RAD counter, available from www.wiki.ed.ac.uk/display/RADSequencing/Home ; last accessed October 15, 2015, we have estimated that 60 individually barcoded samples can be pooled together when using the SbfI restriction enzyme (New England Biolabs). .. By using an average genome size in the target group of 1C = 1.9 pg ( ) and the RAD counter, available from www.wiki.ed.ac.uk/display/RADSequencing/Home ; last accessed October 15, 2015, we have estimated that 60 individually barcoded samples can be pooled together when using the SbfI restriction enzyme (New England Biolabs).

    Plasmid Preparation:

    Article Title: Efficient Production of Papillomavirus Gene Delivery Vectors in Defined In Vitro Reactions
    Article Snippet: All pVITRO constructs were a kind gift from Richard Roden (Department of Pathology, The Johns Hopkins University) and have been described previously by Kwak et al. .. All plasmids were produced in competent Escherichia coli DH5α (BIO-85026, Bioline) and purified using the Plasmid Plus Midi Kit (12945, QIAGEN). pfwB was linearized using SbfI (R3642, New England Biolabs) and pCLucf with XmnI (R0194, New England Biolabs). pCLucf was digested with EcoRI (R3101, New England Biolabs) for the linear split virus used in . .. Digestion was confirmed by agarose gel electrophoresis, and enzymes were heat-inactivated according to the manufacturer’s instructions before use.

    Article Title: Novel System for Efficient Isolation of Clostridium Double-Crossover Allelic Exchange Mutants Enabling Markerless Chromosomal Gene Deletions and DNA Integration
    Article Snippet: Next, the full-length fragment, including the promoter and terminator, was PCR amplified from p94FDH and cloned into the NotI (NEB)-linearized and blunt-ended pKOSIGK_mazF plasmid, yielding plasmid pKISIGK::FDH_mazF. .. The p94FLP_asRepL plasmid was constructed by treating the p94FLP plasmid with SbfI (NEB) to linearize it and subsequently clone the bgaR -asRepL fragment, yielding plasmid p94FLP_asRepL. .. The lactose-inducible promoter and its divergent regulator ( bgaR -PbgaL ) were utilized and fused immediately upstream of a 100-bp antisense RNA (asRNA) that targets the first 100 bp of the repL transcript coding for the replication protein (RepL), creating plasmid p94FLP_asRepL.

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: The mammalian codon optimized GluClβ receptor from C. elegans ( ) was subcloned into the pGEMhe vector and the stop codon of the receptor was mutated from TAG to TGA. .. The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion).

    Article Title: Expression and Function of Aminopeptidase N/CD13 Produced by Fibroblast Like Synoviocytes in Rheumatoid Arthritis: Role of CD13 in Chemotaxis of Cytokine Activated T cells Independent of Enzymatic Activity
    Article Snippet: An E355Q enzymatic inactive mutant was created. .. The plasmid (1μg DNA) was linearized by double digestion with SgrAI (2U) and SbfI (10U) (New England Biolabs). .. The 7.3kb fragment was purified from a 1% agarose gel using the QIAquick Gel Extraction Kit (Qiagen).

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: All constructs were in the pGEMhe vector, a cDNA plasmid optimized for protein expression in Xenopus oocytes. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN).

    Article Title: Early Passage Mesenchymal Stem Cells Display Decreased Radiosensitivity and Increased DNA Repair Activity
    Article Snippet: For overexpression of PARP‐1, MSCs were transduced with PLAS2w vector carrying full length PARP‐1 gene from National Yang‐Ming University VYM Genome Research Center. .. In brief, PLAS2w plasmid was digested by SbfI/AgeI restriction enzymes (New England Biolabs; Ipswich MA; https://www.neb.com /) and the full length of PARP‐1 gene was ligased by T7 DNA ligase (New England Biolabs). .. Viral production of the constructed plasmid with lentivirus system was performed by National Science Council RNAi core facility, Academia Sinica, Taiwan.

    Article Title: In Search of Novel Drug Target Sites on Estrogen Receptors Using RNA Aptamers
    Article Snippet: The human ERα coding gene in pSNAP-hERα vector was lifted from pJ3-FLAG-hERα (a gift from Dr. Carolyn L. Smith, Baylor College of Medicine). .. It was cloned immediately downstream of the SNAP coding sequence between the SbfI (5′ end) and XhoI (3′ end) sites in the pSNAP-tag® (m) vector (New England Biolabs). pSUPER.retro.puro was obtained from OligoEngine. pSHAG-MAGIC has been described previously (Paddison et al., ). .. Reporter vectors 3×ERE TATA luc (Addgene plasmid 11354), 2×PRE TK luc (Addgene plasmid 11350), and pGL3 luc (Promega) all contained a firefly ( Photinus pyralis ) luciferase gene.

    Article Title: Phenotype of the Most Common "Slow Acetylator" Arylamine N-Acetyltransferase 1 Genetic Variant (NAT1*14B) Is Substrate-Dependent
    Article Snippet: The NATb /NAT1 * 4 construct was then ligated into pcDNA5/FRT using T4 ligase (New England Biolabs). .. To construct the NATb/ NAT1 * 14B pcDNA5/FRT plasmid, the NATb/ NAT1 * 4 pcDNA5/FRT and a previously constructed NAT1 * 14B allelic construct expressed in a yeast vector, pESP-3 (Agilent Technologies) , were both incubated at 37°C with restriction enzymes SbfI and AflII (New England Biolabs). .. After restriction digestion, the NATb/ NAT1 * 4 pcDNA5/FRT and the 476-base pair segment of NAT1 * 14B (including G560A) were gel purified and ligated using T4 ligase (New England Biolabs).

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. The relative replication capacity of the virus was determined by measuring the amount of p24 antigen produced 72 h after transfection.

    Article Title: Non-Invasive Fluorescent Monitoring of Ovarian Cancer in an Immunocompetent Mouse Model
    Article Snippet: Paragraph title: 4.2. pROSA-puro-iRFP720 Vector Generation ... pROSA26-1 was digested with SbfI/XhoI (NEB, Ipswich, MA, USA) to release a 4434 bp product.

    Article Title: Na+ Permeation and Block of hERG Potassium Channels
    Article Snippet: The final PCR product was cloned into Zero Blunt Vector (Invitrogen). .. A 1,055-bp fragment containing the point mutation was cut by BstEII and SbfI (New England Biolabs, Inc.), and then subcloned into the pcDNA3 vector containing the wild-type (WT) hERG at the same sites ( ). .. All mutations were verified using a high-throughput 48 capillary ABI 3730 sequencer (UCDNA Services, University of Calgary). hERG mutant channels were transiently expressed in HEK 293 cells (American Type Culture Collection).

    Irradiation:

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion). .. Acylation of tRNA was confirmed using MALDI mass spectrometry using a 3-hydroxypicolinic acid matrix.

    Multiplex Assay:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB). .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Selection:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB). .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Agarose Gel Electrophoresis:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop) and quality assessed by agarose gel electrophoresis, and was finally diluted to a concentration of 50 ng/µL in 5 mmol/L Tris, pH 8.5. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: Genomic DNA from each sample was quantified using spectrophotometry (Qubit 2.0 Fluorometer, Invitrogen) and checked for genomic integrity by 0.8% agarose gel electrophoresis. .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop), and its quality was assessed by agarose gel electrophoresis, before being diluted to a concentration of 20 ng/μL [measured by Qubit Fluorometer (Invitrogen)] in 5 mmol/L Tris, pH 8.5. .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Article Title: Amplification Biases and Consistent Recovery of Loci in a Double-Digest RAD-seq Protocol
    Article Snippet: We then double-digest ∼1.0 µg of DNA with high fidelity versions of the SbfI and EcoRI restriction enzymes (New England Biolabs); when less DNA is available, we have had good success starting with as little as 0.17 µg of genomic DNA. .. We then double-digest ∼1.0 µg of DNA with high fidelity versions of the SbfI and EcoRI restriction enzymes (New England Biolabs); when less DNA is available, we have had good success starting with as little as 0.17 µg of genomic DNA.

    In Vitro:

    Article Title: Secondary Ammonium Agonists Make Dual Cation-π Interactions in α4β2 Nicotinic Receptors
    Article Snippet: Site-directed mutagenesis was performed by PCR using the Stratagene QuikChange protocol, and primers ordered from Integrated DNA Technologies. .. Circular cDNA was linearized with SbfI (New England Biolabs) and then transcribed in vitro using T7 mMessage mMachine kit (Life Technologies), with a purification step after each process (QIAGEN). .. Final concentrations were quantified by UV spectroscopy.

    Next-Generation Sequencing:

    Article Title: Genomic Changes Associated with Reproductive and Migratory Ecotypes in Sockeye Salmon (Oncorhynchus nerka)
    Article Snippet: For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.). .. For each individual sample, 500 ng of DNA was digested using the SbfI restriction enzyme (New England BioLabs Inc.).

    Spectrophotometry:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop) and quality assessed by agarose gel electrophoresis, and was finally diluted to a concentration of 50 ng/µL in 5 mmol/L Tris, pH 8.5. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Article Title: A High Density Genetic Map Derived from RAD Sequencing and Its Application in QTL Analysis of Yield-Related Traits in Vigna unguiculata
    Article Snippet: Genomic DNA from each sample was quantified using spectrophotometry (Qubit 2.0 Fluorometer, Invitrogen) and checked for genomic integrity by 0.8% agarose gel electrophoresis. .. The general protocol was as follows: first, each sample of genomic DNA (~0.5 μg per sample) from the 168 individuals and their two parents was digested with 20 units (U) of EcoR I or Sbf I (New England Biolabs, NEB) for 15 min at 37°C in a 50-μL (microlitre) reaction and then heat denatured at 65°C.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop), and its quality was assessed by agarose gel electrophoresis, before being diluted to a concentration of 20 ng/μL [measured by Qubit Fluorometer (Invitrogen)] in 5 mmol/L Tris, pH 8.5. .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Produced:

    Article Title: Efficient Production of Papillomavirus Gene Delivery Vectors in Defined In Vitro Reactions
    Article Snippet: All pVITRO constructs were a kind gift from Richard Roden (Department of Pathology, The Johns Hopkins University) and have been described previously by Kwak et al. .. All plasmids were produced in competent Escherichia coli DH5α (BIO-85026, Bioline) and purified using the Plasmid Plus Midi Kit (12945, QIAGEN). pfwB was linearized using SbfI (R3642, New England Biolabs) and pCLucf with XmnI (R0194, New England Biolabs). pCLucf was digested with EcoRI (R3101, New England Biolabs) for the linear split virus used in . .. Digestion was confirmed by agarose gel electrophoresis, and enzymes were heat-inactivated according to the manufacturer’s instructions before use.

    Article Title: Functional Evaluation of Key Interactions Evident in the Structure of the Eukaryotic Cys-Loop Receptor GluCl
    Article Snippet: Mutagenesis of the GluClβ receptor was performed using the QuikChange protocol (Stratagene), and for nonsense suppression experiments, a TAG codon was mutated into the site of interest. .. The cDNA was linearized using SbfI (New England Biolabs) and mRNA was produced from the linearized plasmids by using the T7 mMessage Machine kit (Ambion). .. 74mer THG73 tRNA was in vitro transcribed from a DNA oligonucleotide template containing two 5′ methoxy (C2′ position) nucleotides to site-specifically truncate transcription by the T7 MEGAshortscript kit (Ambion).

    Article Title: Changes in codon-pair bias of human immunodeficiency virus type 1 have profound effects on virus replication in cell culture
    Article Snippet: Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites. .. Protease sequences were PCR amplified with oligonuclotides Apa1988 (5′-ACATAGCCAAAAATTGCAGGGCCCCTAG-3′) and Sbf2838 (5′-CTGATTTTTTCTGTTTTAACCCTGCAGGATG-3′), digested with Apa I and Sbf I (New England Biolabs), and cloned into pNL4-3-deltaE-EGFP between the Apa I and Sbf I sites.

    Concentration Assay:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: The RAD specific P1 and P2 paired-end adapters and library amplification PCR primer sequences used in this study are detailed in Baxter et al. . .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume. .. The reactions (12 µL final volumes) were then heat inactivated at 65°C for 20 minutes.

    Article Title: Accuracy of Genomic Evaluations of Juvenile Growth Rate in Common Carp (Cyprinus carpio) Using Genotyping by Sequencing
    Article Snippet: Each sample was quantified by spectrophotometry (Nanodrop), and its quality was assessed by agarose gel electrophoresis, before being diluted to a concentration of 20 ng/μL [measured by Qubit Fluorometer (Invitrogen)] in 5 mmol/L Tris, pH 8.5. .. Briefly, each sample (0.72 μg parental DNA/0.24 μg offspring DNA) was digested at 37°C for 60 min with Sbf I (recognizing the CCTGCA| GG motif) high fidelity restriction enzyme (New England Biolabs, NEB).

    Fluorescence In Situ Hybridization:

    Article Title: Mapping and Validation of the Major Sex-Determining Region in Nile Tilapia (Oreochromis niloticus L.) Using RAD Sequencing
    Article Snippet: DNA was extracted from fin tissue of the fish using the REALPure genomic DNA extraction kit (Durviz S.L.) and treated with RNase to remove residual RNA from the sample. .. Each sample (0.72 µg parental DNA/0.24 µg offspring DNA) was digested at 37°C for 40 minutes with Sbf I (recognising the CCTGCA|GG motif) high fidelity restriction enzyme (New England Biolabs; NEB) using 6 U Sbf I per µg genomic DNA in 1× Reaction Buffer 4 (NEB) at a final concentration of c. 1 µg DNA per 50 µL reaction volume.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • N/A
    SbfI HF 2 500 units
      Buy from Supplier

    99
    New England Biolabs sbfi
    Sbfi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sbfi/product/New England Biolabs
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    sbfi - by Bioz Stars, 2019-07
    99/100 stars
      Buy from Supplier

    Image Search Results