bstni  (New England Biolabs)


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    BstNI
    Description:
    BstNI 15 000 units
    Catalog Number:
    r0168l
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    249
    Category:
    Restriction Enzymes
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    15 000 units
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    New England Biolabs bstni
    BstNI
    BstNI 15 000 units
    https://www.bioz.com/result/bstni/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bstni - by Bioz Stars, 2021-03
    93/100 stars

    Images

    1) Product Images from "Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome"

    Article Title: Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome

    Journal: Nucleic Acids Research

    doi:

    Flow diagram of rare point-mutational analysis by CDCE/hifiPCR: natural versus ligated clamp. The copy numbers of the wild-type and of a mutant added at an initial fraction of 5 × 10 –5 are shown after each step. Restriction digestion by BstNI and DraI liberates the HPRT target-embedded fragment of 438 bp from genomic DNA. The lines in this fragment indicate the positions of the G to A transition and of the G to T transversion carried by the internal standards of the 438 bp PCR fragment and of the HPRT Munich cells, respectively. The restriction-recognition sites of AhdI, ApoI and HinfI are also indicated by the lines. The open and filled bars indicate the positions of the probes (Probe 1 and Probe 2) used for target isolation and of the PCR primers (P3 and P1), respectively.
    Figure Legend Snippet: Flow diagram of rare point-mutational analysis by CDCE/hifiPCR: natural versus ligated clamp. The copy numbers of the wild-type and of a mutant added at an initial fraction of 5 × 10 –5 are shown after each step. Restriction digestion by BstNI and DraI liberates the HPRT target-embedded fragment of 438 bp from genomic DNA. The lines in this fragment indicate the positions of the G to A transition and of the G to T transversion carried by the internal standards of the 438 bp PCR fragment and of the HPRT Munich cells, respectively. The restriction-recognition sites of AhdI, ApoI and HinfI are also indicated by the lines. The open and filled bars indicate the positions of the probes (Probe 1 and Probe 2) used for target isolation and of the PCR primers (P3 and P1), respectively.

    Techniques Used: Flow Cytometry, Mutagenesis, Polymerase Chain Reaction, Isolation

    2) Product Images from "Development and Evaluation of a Real-Time PCR Assay for Detection of Klebsiella pneumoniae Carbapenemase Genes ▿"

    Article Title: Development and Evaluation of a Real-Time PCR Assay for Detection of Klebsiella pneumoniae Carbapenemase Genes ▿

    Journal:

    doi: 10.1128/JCM.01550-08

    Real-time PCR assay for bla KPC . Real-time PCR using a TaqMan probe generates a 399-bp amplicon for all bla KPC genes. Amplicons from positive samples can be digested with BstNI and RsaI to detect the nucleotide polymorphisms in the PCR amplicon reported
    Figure Legend Snippet: Real-time PCR assay for bla KPC . Real-time PCR using a TaqMan probe generates a 399-bp amplicon for all bla KPC genes. Amplicons from positive samples can be digested with BstNI and RsaI to detect the nucleotide polymorphisms in the PCR amplicon reported

    Techniques Used: Real-time Polymerase Chain Reaction, Amplification, Polymerase Chain Reaction

    3) Product Images from "A Mutation in Hnrnph1 That Decreases Methamphetamine-Induced Reinforcement, Reward, and Dopamine Release and Increases Synaptosomal hnRNP H and Mitochondrial Proteins"

    Article Title: A Mutation in Hnrnph1 That Decreases Methamphetamine-Induced Reinforcement, Reward, and Dopamine Release and Increases Synaptosomal hnRNP H and Mitochondrial Proteins

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1808-19.2019

    Hnrnph1 whole-brain mRNA and hnRNP H whole-body protein expression in WT, H1 +/− , and H1 −/− mice. H1 −/− mice and WT littermates were generated by intercrossing H1 +/− and H1 +/− . Embryos were harvested at E12 for genotyping using a restriction enzyme-based assay. A , A PCR amplicon capturing the deleted region was digested with BstNI. WT mice had two copies of two BstNI restriction sites; thus, restriction digest produced t hree fragments (58, 157, and 153 bp) corresponding to two bands on the gel. H1 −/− mice had two copies of a single BstNI restriction site; thus, restriction digest produced two fragments (153 and 198 bp). H1 +/− mice possessed one copy of each of the two BstNI restriction sites and one copy of a single BstNI restriction site; thus, restriction digest produced 5 fragments (58, 153, 153, 157, and 198 bp) corresponding to three bands on the gel. B , There was a gene dosage-dependent increase in the transcript level Hnrnph1 in H1 +/− and H1 −/− mice. The 1.5 increase in Hnnph1 transcript level in H1 +/− ). The > 2-fold increase in Hnrnph1 transcript level in H1 −/− with two copies of the mutation provides further functional support for increased expression of the mutant transcript. * p = 0.031 (unpaired t test for WT vs H1 −/− ). C , There was no genotypic difference in Hnrnph2 transcript level. D–G , Protein expression of hnRNP H in WT, H1 +/− , and H1 −/− mice. There was no significant genotypic difference in hnRNP H protein expression using an antibody targeting the C terminus of hnRNP H ( D , E ) or the N terminus of hnRNP H ( F , G ). n = 4 per genotype.
    Figure Legend Snippet: Hnrnph1 whole-brain mRNA and hnRNP H whole-body protein expression in WT, H1 +/− , and H1 −/− mice. H1 −/− mice and WT littermates were generated by intercrossing H1 +/− and H1 +/− . Embryos were harvested at E12 for genotyping using a restriction enzyme-based assay. A , A PCR amplicon capturing the deleted region was digested with BstNI. WT mice had two copies of two BstNI restriction sites; thus, restriction digest produced t hree fragments (58, 157, and 153 bp) corresponding to two bands on the gel. H1 −/− mice had two copies of a single BstNI restriction site; thus, restriction digest produced two fragments (153 and 198 bp). H1 +/− mice possessed one copy of each of the two BstNI restriction sites and one copy of a single BstNI restriction site; thus, restriction digest produced 5 fragments (58, 153, 153, 157, and 198 bp) corresponding to three bands on the gel. B , There was a gene dosage-dependent increase in the transcript level Hnrnph1 in H1 +/− and H1 −/− mice. The 1.5 increase in Hnnph1 transcript level in H1 +/− ). The > 2-fold increase in Hnrnph1 transcript level in H1 −/− with two copies of the mutation provides further functional support for increased expression of the mutant transcript. * p = 0.031 (unpaired t test for WT vs H1 −/− ). C , There was no genotypic difference in Hnrnph2 transcript level. D–G , Protein expression of hnRNP H in WT, H1 +/− , and H1 −/− mice. There was no significant genotypic difference in hnRNP H protein expression using an antibody targeting the C terminus of hnRNP H ( D , E ) or the N terminus of hnRNP H ( F , G ). n = 4 per genotype.

    Techniques Used: Expressing, Mouse Assay, Generated, Enzymatic Assay, Polymerase Chain Reaction, Amplification, Produced, Mutagenesis, Functional Assay

    4) Product Images from "Selection of functional human antibodies from retroviral display libraries"

    Article Title: Selection of functional human antibodies from retroviral display libraries

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gni033

    Fingerprint analysis of the selected scFvs. RT–PCR fragments derived from genomic viral RNA representing the scFv fragment coding sequences before ( A ) and after three rounds of selection ( B ) were cloned into bacteria. Colony-PCR fragments from 50 randomly picked bacterial clones were restricted with BstNI, respectively. Similar restriction patterns were grouped and the patterns of the most prominent clones L9, L28 and L6 are indicated.
    Figure Legend Snippet: Fingerprint analysis of the selected scFvs. RT–PCR fragments derived from genomic viral RNA representing the scFv fragment coding sequences before ( A ) and after three rounds of selection ( B ) were cloned into bacteria. Colony-PCR fragments from 50 randomly picked bacterial clones were restricted with BstNI, respectively. Similar restriction patterns were grouped and the patterns of the most prominent clones L9, L28 and L6 are indicated.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Selection, Clone Assay, Polymerase Chain Reaction

    5) Product Images from "Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica"

    Article Title: Diagnostic Molecular Markers for Phosphine Resistance in U.S. Populations of Tribolium castaneum and Rhyzopertha dominica

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0121343

    Restriction enzyme digests of PCR products amplified from genomic DNA coding for the DLD gene. M = size marker (100bp DNA Ladder, New England BioLabs Inc., MA USA); lanes 1, 2 and 3 on each gel are digests of individual beetles as noted. Gels A and B are digested products from homozygous resistant (RR), susceptible (SS) or heterozygous (RS) T . castaneum from the TcOK-G population digested with either MboI or BstNI, respectively. Gels C and D are similarly digested products from R . dominica . See text for details.
    Figure Legend Snippet: Restriction enzyme digests of PCR products amplified from genomic DNA coding for the DLD gene. M = size marker (100bp DNA Ladder, New England BioLabs Inc., MA USA); lanes 1, 2 and 3 on each gel are digests of individual beetles as noted. Gels A and B are digested products from homozygous resistant (RR), susceptible (SS) or heterozygous (RS) T . castaneum from the TcOK-G population digested with either MboI or BstNI, respectively. Gels C and D are similarly digested products from R . dominica . See text for details.

    Techniques Used: Polymerase Chain Reaction, Amplification, Marker

    6) Product Images from "Comparative analysis of the end-joining activity of several DNA ligases"

    Article Title: Comparative analysis of the end-joining activity of several DNA ligases

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0190062

    Wild type DNA ligase λ DNA digest ligation assay. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1 ), NruI (G/C Blunt, 2 ), BstNI (5′ SBO, 3 ), Hpy188I (3′SBO, 4 ), NdeI (2 BO, 5 ) and BamHI (4 BO, 6 ), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in the presence of T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl2, 1 mM DTT, 1 mM ATP, 6% polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and, hLig3 (D), respectively. E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.
    Figure Legend Snippet: Wild type DNA ligase λ DNA digest ligation assay. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1 ), NruI (G/C Blunt, 2 ), BstNI (5′ SBO, 3 ), Hpy188I (3′SBO, 4 ), NdeI (2 BO, 5 ) and BamHI (4 BO, 6 ), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in the presence of T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl2, 1 mM DTT, 1 mM ATP, 6% polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with T4 DNA ligase (A), T3 DNA ligase (B), PBCV1 DNA ligase (C) and, hLig3 (D), respectively. E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.

    Techniques Used: Ligation, Agarose Gel Electrophoresis, Staining

    Effect of DBDs on blunt/cohesive end λ DNA Re-ligation. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1), NruI (G/C Blunt, 2), BstNI (5′ SBO, 3), Hpy188I (3′SBO, 4), NdeI (2 BO, 5) and BamHI (4 BO, 6), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with PBCV1-Nterm-Sso7d (A), PBCV1-Cterm-Sso7d terminus (B), PBCV1-Nterm-ZnF (C), PBCV1-Nterm-T4NTD (D). (E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.
    Figure Legend Snippet: Effect of DBDs on blunt/cohesive end λ DNA Re-ligation. Agarose gel electrophoresis of λ DNA cut by EcoRV (A/T Blunt, 1), NruI (G/C Blunt, 2), BstNI (5′ SBO, 3), Hpy188I (3′SBO, 4), NdeI (2 BO, 5) and BamHI (4 BO, 6), generating DNA fragments with ligatable ends. 0.5 ng of the cut DNA was ligated in T4 ligase reaction buffer (50 mM Tris-HCl pH 7.5 @ 25°C, 1 mM ATP and 10 mM MgCl 2 ) or NEBNext ® Quick Ligation reaction buffer (66 mM Tris pH 7.6 @ 25°C, 10 mM MgCl 2 , 1 mM DTT, 1 mM ATP, 6% Polyethylene glycol (PEG 6000)) and 7 μM of the indicated DNA ligase for 1 hour at 25°C. Ligation assays performed with PBCV1-Nterm-Sso7d (A), PBCV1-Cterm-Sso7d terminus (B), PBCV1-Nterm-ZnF (C), PBCV1-Nterm-T4NTD (D). (E) Gel of restriction enzyme digested λ DNA samples as well as a schematic depiction of each substrate. The DNA fragments were visualized using ethidium bromide stain.

    Techniques Used: Ligation, Agarose Gel Electrophoresis, Staining

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Selection of Recombinant Antibodies Specific for Pathogenic Streptococcus suis by Subtractive Phage Display
    Article Snippet: PCR products were analyzed on a 1.5% agarose gel containing 0.5 μg of ethidium bromide ml−1 . .. PCR products were digested with Bst NI (New England Biolabs, Beverly, Mass.) or Ava II (Promega, Madison, Wis.). .. The restriction mixture, containing 50 mM NaCl, 10 mM Tris-HCl (pH 7.9), 10 mM MgCl2 , 1 mM dithiothreitol, and 4 U of Bst NI or Ava II, was added to the PCR mixture in a 1:1 ratio.

    Article Title: A Mutation in Hnrnph1 That Decreases Methamphetamine-Induced Reinforcement, Reward, and Dopamine Release and Increases Synaptosomal hnRNP H and Mitochondrial Proteins
    Article Snippet: .. Genomic DNA was used to amplify a 204 bp PCR product using DreamTaq Green PCR Mastermix (Thermo Fisher Scientific) followed by overnight restriction enzyme digest with BstNI (New England Biolabs). .. Hnrnph1 and Hnrnph2 qRT-PCR for mouse embryo tissue Oligo-dT primers were used to synthesize cDNA from total RNA to examine mRNA expression and qPCR for evaluating gene expression performed using TaqMan SYBR Green (Thermo Fisher Scientific, catalog #4309155).

    Article Title: Genetic diversity among Plasmodium vivax isolates along the Thai–Myanmar border of Thailand
    Article Snippet: One microlitres of template DNA was used to initiate amplification, followed by a second amplification, in which 1 µL of PCR products from the first amplification was used. .. Subsequently, 20 µL of the second PCR products were separately digested by restriction enzymes Alu I and Bst NI (New England Biolabs Inc., UK) according to the manufacture specification for 2 h in a total volume of 30 µL. .. The DNA fragments were separated by electrophoresis on 3 % agarose gel and visualized under UV illumination after ethidium bromide staining.

    Article Title: Rapid Concerted Evolution of Nuclear Ribosomal DNA in Two Tragopogon Allopolyploids of Recent and Recurrent Origin
    Article Snippet: For PCR (50 μl) amplification we used 100 ng of input RNA or 100 ng of genomic DNA as template, 40 pmol of each primer (18Sfor and 5.8Srev), 12 nmol of each dNTP, and 1.9 units of DyNAzyme II DNA polymerase (FINNZYMES, Espoo, Finland). .. Cycling conditions were as follows: initial denaturation step (92°, 180 sec) and 35 cycles of 92° for 20 sec, 57° for 30 sec, and 72° for 30 sec, followed by a final 72° extension for 10 min. To discriminate among ITS-1 variants, the PCR products were digested with Bst NI (New England Biolabs, Beverly, MA), which produces species-specific DNA fragments for each diploid progenitor ( ); the resulting fragments were separated on a 7% polyacrylamide gel. .. The gel was stained after electrophoresis with ethidium bromide and photographed.

    Antiviral Assay:

    Article Title: Selection of Recombinant Antibodies Specific for Pathogenic Streptococcus suis by Subtractive Phage Display
    Article Snippet: PCR products were analyzed on a 1.5% agarose gel containing 0.5 μg of ethidium bromide ml−1 . .. PCR products were digested with Bst NI (New England Biolabs, Beverly, Mass.) or Ava II (Promega, Madison, Wis.). .. The restriction mixture, containing 50 mM NaCl, 10 mM Tris-HCl (pH 7.9), 10 mM MgCl2 , 1 mM dithiothreitol, and 4 U of Bst NI or Ava II, was added to the PCR mixture in a 1:1 ratio.

    Nucleic Acid Electrophoresis:

    Article Title: Sequential, Divergent, and Cooperative Requirements of Foxl2a and Foxl2b in Ovary Development and Maintenance of Zebrafish
    Article Snippet: The gDNA fragments including the target site of the TALENs were amplified using foxl2a and foxl2b gene-specific primers ( ). .. After digestion with Bst N I (restriction enzyme cutting site in the spacer of foxl2a ) or Bsr D I (restriction enzyme cutting site in the spacer of foxl2b ) (New England Biolabs, Beverly, MA) respectively at 60° and 65° for 2 hr, the uncleaved DNA fragments were separated by gel electrophoresis and cloned into pMD-18T vector (Promega). .. Sequence alignments were generated to analyze whether or not the gDNA fragments from embryos injected with TALEN mRNAs were mutated.

    Clone Assay:

    Article Title: Sequential, Divergent, and Cooperative Requirements of Foxl2a and Foxl2b in Ovary Development and Maintenance of Zebrafish
    Article Snippet: The gDNA fragments including the target site of the TALENs were amplified using foxl2a and foxl2b gene-specific primers ( ). .. After digestion with Bst N I (restriction enzyme cutting site in the spacer of foxl2a ) or Bsr D I (restriction enzyme cutting site in the spacer of foxl2b ) (New England Biolabs, Beverly, MA) respectively at 60° and 65° for 2 hr, the uncleaved DNA fragments were separated by gel electrophoresis and cloned into pMD-18T vector (Promega). .. Sequence alignments were generated to analyze whether or not the gDNA fragments from embryos injected with TALEN mRNAs were mutated.

    Plasmid Preparation:

    Article Title: Sequential, Divergent, and Cooperative Requirements of Foxl2a and Foxl2b in Ovary Development and Maintenance of Zebrafish
    Article Snippet: The gDNA fragments including the target site of the TALENs were amplified using foxl2a and foxl2b gene-specific primers ( ). .. After digestion with Bst N I (restriction enzyme cutting site in the spacer of foxl2a ) or Bsr D I (restriction enzyme cutting site in the spacer of foxl2b ) (New England Biolabs, Beverly, MA) respectively at 60° and 65° for 2 hr, the uncleaved DNA fragments were separated by gel electrophoresis and cloned into pMD-18T vector (Promega). .. Sequence alignments were generated to analyze whether or not the gDNA fragments from embryos injected with TALEN mRNAs were mutated.

    Agarose Gel Electrophoresis:

    Article Title: Development and Evaluation of a Real-Time PCR Assay for Detection of Klebsiella pneumoniae Carbapenemase Genes ▿
    Article Snippet: Amplification was performed using recombinant Taq Polymerase (Qiagen) at a final magnesium concentration of 2 mM. .. The following cycling conditions were used: 95o for 2 min, followed by 35 cycles of 94°C for 2 s, 62°C for 10 s, and 72°C for 15 s. The amplicons from KPC-positive samples were then digested at 48°C for 1 h in RsaI and BstNI (New England Biolabs, Ipswitch, MA) using NEB buffer 2 (New England Biolabs) and electrophoresed on a 2% agarose gel to differentiate bla KPC-1 , bla KPC-2 , and bla KPC-3 . ..

    Isolation:

    Article Title: Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome
    Article Snippet: A DNA yield of > 90% and a ratio of A 260 to A 280 in the range of 1.6–2.0 were estimated by a UV spectrophotometer. .. Restriction digestion by BstNI and DraI (New England Biolabs, Beverly, MA) was performed with the samples of genomic DNA to liberate the HPRT target-embedded fragment of 438 bp suitable for target isolation by probe-target hybridization coupled with a biotin–streptavidin capture system ( ). ..

    Hybridization:

    Article Title: Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome
    Article Snippet: A DNA yield of > 90% and a ratio of A 260 to A 280 in the range of 1.6–2.0 were estimated by a UV spectrophotometer. .. Restriction digestion by BstNI and DraI (New England Biolabs, Beverly, MA) was performed with the samples of genomic DNA to liberate the HPRT target-embedded fragment of 438 bp suitable for target isolation by probe-target hybridization coupled with a biotin–streptavidin capture system ( ). ..

    Size-exclusion Chromatography:

    Article Title: Rapid Concerted Evolution of Nuclear Ribosomal DNA in Two Tragopogon Allopolyploids of Recent and Recurrent Origin
    Article Snippet: For PCR (50 μl) amplification we used 100 ng of input RNA or 100 ng of genomic DNA as template, 40 pmol of each primer (18Sfor and 5.8Srev), 12 nmol of each dNTP, and 1.9 units of DyNAzyme II DNA polymerase (FINNZYMES, Espoo, Finland). .. Cycling conditions were as follows: initial denaturation step (92°, 180 sec) and 35 cycles of 92° for 20 sec, 57° for 30 sec, and 72° for 30 sec, followed by a final 72° extension for 10 min. To discriminate among ITS-1 variants, the PCR products were digested with Bst NI (New England Biolabs, Beverly, MA), which produces species-specific DNA fragments for each diploid progenitor ( ); the resulting fragments were separated on a 7% polyacrylamide gel. .. The gel was stained after electrophoresis with ethidium bromide and photographed.

    Incubation:

    Article Title: G388R mutation of the FGFR4 gene is not relevant to breast cancer prognosis
    Article Snippet: The digestion reactions contained 10 μ l of PCR product, 0.5 μ l of Bst NI (5 U; New England Biolabs, Beverly, MA, USA), 2 μ l of 10 × NEBuffer 2 (supplied with the enzyme) and 0.2 μ l bovine serum albumin (100 mg l−1 ) in a final volume of 20 μ l. These components were incubated for 60 min at 60°C. .. The digestion reactions contained 10 μ l of PCR product, 0.5 μ l of Bst NI (5 U; New England Biolabs, Beverly, MA, USA), 2 μ l of 10 × NEBuffer 2 (supplied with the enzyme) and 0.2 μ l bovine serum albumin (100 mg l−1 ) in a final volume of 20 μ l. These components were incubated for 60 min at 60°C. .. After the reaction ended, 10 μ l of the PCR mixture were mixed with a loading buffer and electrophoresed in a 4% agarose 1000® gel (Life Technologies, Carlsbad, CA, USA).

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    New England Biolabs bstni
    Flow diagram of rare point-mutational analysis by CDCE/hifiPCR: natural versus ligated clamp. The copy numbers of the wild-type and of a mutant added at an initial fraction of 5 × 10 –5 are shown after each step. Restriction digestion by <t>BstNI</t> and <t>DraI</t> liberates the HPRT target-embedded fragment of 438 bp from genomic DNA. The lines in this fragment indicate the positions of the G to A transition and of the G to T transversion carried by the internal standards of the 438 bp PCR fragment and of the HPRT Munich cells, respectively. The restriction-recognition sites of AhdI, ApoI and HinfI are also indicated by the lines. The open and filled bars indicate the positions of the probes (Probe 1 and Probe 2) used for target isolation and of the PCR primers (P3 and P1), respectively.
    Bstni, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bstni/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bstni - by Bioz Stars, 2021-03
    93/100 stars
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    Flow diagram of rare point-mutational analysis by CDCE/hifiPCR: natural versus ligated clamp. The copy numbers of the wild-type and of a mutant added at an initial fraction of 5 × 10 –5 are shown after each step. Restriction digestion by BstNI and DraI liberates the HPRT target-embedded fragment of 438 bp from genomic DNA. The lines in this fragment indicate the positions of the G to A transition and of the G to T transversion carried by the internal standards of the 438 bp PCR fragment and of the HPRT Munich cells, respectively. The restriction-recognition sites of AhdI, ApoI and HinfI are also indicated by the lines. The open and filled bars indicate the positions of the probes (Probe 1 and Probe 2) used for target isolation and of the PCR primers (P3 and P1), respectively.

    Journal: Nucleic Acids Research

    Article Title: Ligation of high-melting-temperature 'clamp' sequence extends the scanning range of rare point-mutational analysis by constant denaturant capillary electrophoresis (CDCE) to most of the human genome

    doi:

    Figure Lengend Snippet: Flow diagram of rare point-mutational analysis by CDCE/hifiPCR: natural versus ligated clamp. The copy numbers of the wild-type and of a mutant added at an initial fraction of 5 × 10 –5 are shown after each step. Restriction digestion by BstNI and DraI liberates the HPRT target-embedded fragment of 438 bp from genomic DNA. The lines in this fragment indicate the positions of the G to A transition and of the G to T transversion carried by the internal standards of the 438 bp PCR fragment and of the HPRT Munich cells, respectively. The restriction-recognition sites of AhdI, ApoI and HinfI are also indicated by the lines. The open and filled bars indicate the positions of the probes (Probe 1 and Probe 2) used for target isolation and of the PCR primers (P3 and P1), respectively.

    Article Snippet: Restriction digestion by BstNI and DraI (New England Biolabs, Beverly, MA) was performed with the samples of genomic DNA to liberate the HPRT target-embedded fragment of 438 bp suitable for target isolation by probe-target hybridization coupled with a biotin–streptavidin capture system ( ).

    Techniques: Flow Cytometry, Mutagenesis, Polymerase Chain Reaction, Isolation

    Establishment of  foxl2a  and  foxl2b  knockout mutant lines by TALEN. (A and B) The TALEN target sites of zebrafish (A)  foxl2a  and (B)  foxl2b . The coding and untranslated exon regions are depicted as solid and open boxes, respectively. The left and right TALEN binding sites are indicated by underlining. Cleavage sites with  Bst N I and  Bsr D I in the spacer are shown by blue color, and forward and reverse primers (F primer and R primer) are indicated in the corresponding sites. (C and D) Detection of (C)  foxl2a  and (D)  foxl2b  mutants by  Bst N I or  Bsr D I digestion. The amplified fragment sizes (bp) are shown on the right. (E and F) Sequences of different indels of TALEN-induced (E)  foxl2a  and (F)  foxl2b  mutants in F 0  embryos. A total of eight and six indels (number of embryos are indicated in each bracket) at targeted locus are shown for (E)  foxl2a  and (F)  foxl2b , and the numbers at the right-hand side indicate the number of deleted base pairs. (G) Transcription-level confirmation of  foxl2a  and  foxl2b  mutants by RT-PCR. The detected primers are shown on the left, and primer P2 is specific for the deleted sequences. The  actb1  was used as control. F, forward; R, reverse; M, marker.

    Journal: Genetics

    Article Title: Sequential, Divergent, and Cooperative Requirements of Foxl2a and Foxl2b in Ovary Development and Maintenance of Zebrafish

    doi: 10.1534/genetics.116.199133

    Figure Lengend Snippet: Establishment of foxl2a and foxl2b knockout mutant lines by TALEN. (A and B) The TALEN target sites of zebrafish (A) foxl2a and (B) foxl2b . The coding and untranslated exon regions are depicted as solid and open boxes, respectively. The left and right TALEN binding sites are indicated by underlining. Cleavage sites with Bst N I and Bsr D I in the spacer are shown by blue color, and forward and reverse primers (F primer and R primer) are indicated in the corresponding sites. (C and D) Detection of (C) foxl2a and (D) foxl2b mutants by Bst N I or Bsr D I digestion. The amplified fragment sizes (bp) are shown on the right. (E and F) Sequences of different indels of TALEN-induced (E) foxl2a and (F) foxl2b mutants in F 0 embryos. A total of eight and six indels (number of embryos are indicated in each bracket) at targeted locus are shown for (E) foxl2a and (F) foxl2b , and the numbers at the right-hand side indicate the number of deleted base pairs. (G) Transcription-level confirmation of foxl2a and foxl2b mutants by RT-PCR. The detected primers are shown on the left, and primer P2 is specific for the deleted sequences. The actb1 was used as control. F, forward; R, reverse; M, marker.

    Article Snippet: After digestion with Bst N I (restriction enzyme cutting site in the spacer of foxl2a ) or Bsr D I (restriction enzyme cutting site in the spacer of foxl2b ) (New England Biolabs, Beverly, MA) respectively at 60° and 65° for 2 hr, the uncleaved DNA fragments were separated by gel electrophoresis and cloned into pMD-18T vector (Promega).

    Techniques: Knock-Out, Mutagenesis, Binding Assay, Amplification, Reverse Transcription Polymerase Chain Reaction, Marker

    35S rRNA gene expression in diploid and allotetraploid species of Tragopogon. Primary RNA transcripts were analyzed in the ITS-1 region by RT-PCR using Bst NI restriction site polymorphisms. The digestion products of PCR reactions were separated on a 7%

    Journal: Genetics

    Article Title: Rapid Concerted Evolution of Nuclear Ribosomal DNA in Two Tragopogon Allopolyploids of Recent and Recurrent Origin

    doi: 10.1534/genetics.104.032839

    Figure Lengend Snippet: 35S rRNA gene expression in diploid and allotetraploid species of Tragopogon. Primary RNA transcripts were analyzed in the ITS-1 region by RT-PCR using Bst NI restriction site polymorphisms. The digestion products of PCR reactions were separated on a 7%

    Article Snippet: Cycling conditions were as follows: initial denaturation step (92°, 180 sec) and 35 cycles of 92° for 20 sec, 57° for 30 sec, and 72° for 30 sec, followed by a final 72° extension for 10 min. To discriminate among ITS-1 variants, the PCR products were digested with Bst NI (New England Biolabs, Beverly, MA), which produces species-specific DNA fragments for each diploid progenitor ( ); the resulting fragments were separated on a 7% polyacrylamide gel.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    PCR – RFLP analysis of FGFR4 gene G388R mutation. Lane 1, Hae III-digested pBR322 size marker; lane 2, amplification control; lane 3, wild-type control (Gly/Gly), Bst NI – digested; lane 4, heterozygote carrier (Gly/Arg), Bst NI – digested; lane 5, homozygote carrier (Arg/Arg), Bst NI – digested.

    Journal: British Journal of Cancer

    Article Title: G388R mutation of the FGFR4 gene is not relevant to breast cancer prognosis

    doi: 10.1038/sj.bjc.6601450

    Figure Lengend Snippet: PCR – RFLP analysis of FGFR4 gene G388R mutation. Lane 1, Hae III-digested pBR322 size marker; lane 2, amplification control; lane 3, wild-type control (Gly/Gly), Bst NI – digested; lane 4, heterozygote carrier (Gly/Arg), Bst NI – digested; lane 5, homozygote carrier (Arg/Arg), Bst NI – digested.

    Article Snippet: The digestion reactions contained 10 μ l of PCR product, 0.5 μ l of Bst NI (5 U; New England Biolabs, Beverly, MA, USA), 2 μ l of 10 × NEBuffer 2 (supplied with the enzyme) and 0.2 μ l bovine serum albumin (100 mg l−1 ) in a final volume of 20 μ l. These components were incubated for 60 min at 60°C.

    Techniques: Polymerase Chain Reaction, Mutagenesis, Marker, Amplification