bsp1286i  (New England Biolabs)


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    Name:
    Bsp1286I
    Description:
    Bsp1286I 500 units
    Catalog Number:
    r0120l
    Price:
    65
    Size:
    500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs bsp1286i
    Bsp1286I
    Bsp1286I 500 units
    https://www.bioz.com/result/bsp1286i/product/New England Biolabs
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    bsp1286i - by Bioz Stars, 2020-02
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. .. To identify the nature of indel mutations, PCR products were cloned into pGEM-T easy vector (Promega, #A1360) and DNA was isolated from individual clones and sequenced using an SP6 primer.

    Amplification:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: A 408-bp fragment spanning the miR-430 target site was amplified from wild-type or TALEN-injected zebrafish gDNA using Phusion DNA polymerase (NEB, #M0532) and flanking primers lfty2-Fwd (5′-CCCATGATGTACCTGGTCAAAA-3′) and lfty2-Rev (5′-GCTGTGGTGACCCCTAATGAAT-3′). .. The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C.

    Article Title: Molecular and Clinical Characterization of the Variable Phenotype in Korean Families with Hearing Loss Associated with the Mitochondrial A1555G Mutation
    Article Snippet: .. For the identification of the TRMU gene mutation G28T (A10S), the PCR amplification of exon 1 of the TRMU gene was performed using a previously reported primer, and the PCR products were analyzed using PCR-RFLP with Bsp 1286I (New England Biolabs, Ipswich, MA, USA) . ..

    Article Title: A MITE-based genotyping method to reveal hundreds of DNA polymorphisms in an animal genome after a few generations of artificial selection
    Article Snippet: Preparation of genomic representations For each sample, digestion and ligation reactions were carried out simultaneously at 37°C for 3 hours on 50 ng of genomic DNA, using 2 units of restriction enzyme Bsp1286I (New England Biolabs, NEB), 80 units of T4 DNA ligase (NEB) and 0.05 μM Bsp1286I adaptors I (Table ), in a buffer with final concentrations of 10 mM Tris-OAc, 50 mM KOAc, 10 mM Mg(OAc)2, 5 mM DTT (pH 7.8), 1 mM ATP and 100 ng/ml Bovine Serum Albumin (NEB). .. The obtained ligated products served as template in a first round of PCR amplification.

    Article Title: DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis
    Article Snippet: .. DNA prepared by PCR was amplified with synthetic primers from synthetic templates by using Ready-To-Go PCR Beads (Amersham Biosciences), and the synthetic segments were removed from the final products by restriction digests with Bsp 1286I and Sfc I (NEB). .. The resulting 70-mer (dS70 ) and 78-mer restriction products were separated by denaturing PAGE to generate single-stranded PCR products. dC500 and dA1300 were purchased from Amersham Biosciences.

    Article Title: Genetic Structure, Linkage Disequilibrium and Signature of Selection in Sorghum: Lessons from Physically Anchored DArT Markers
    Article Snippet: Bsp 1286I adapters were ligated to the digested DNA with T4 DNA ligase (NEB). .. A 1-µl aliquot of the ligation product was used as the template in two amplification reactions with one DArT-Bsp 1286I primer ( 5′-GAG TAG TGC CAG AAC GGT C-3′ ) and two MITE (transposable elements) DArT-TIR specific primers ( 5′-TTT TTG GAA CTA AAC AAG GCC-3′ and 5′-G GGT GAA CTA AAC AAG GCC-3′ ).

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR procedure was as follows: an initial denaturation step at 94°C for 2 min and then amplified for 35 cycles at 94°C for 30 sec, 57°C for 40 sec, and 72°C for 40 sec, followed by a final extension step at 72°C for 7 min. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: .. After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. The digested products were analyzed in 1.5% agarose gel stained with ethidium bromide.

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: The T to C transition at position 3111 of the 3’ untranslated region of hClock was amplified using the primers described by Katzenberg et al. . .. Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele.

    Electrophoresis:

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR products were digested with the restriction enzyme Bsp1286 I (SduI), were separated by electrophoresis on 1,5% (w/v) agarose gel, and were visualized by ethidium bromide staining. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele. .. The restriction fragments were resolved by electrophoresis on a 3% agarose gel.

    Incubation:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: Restriction enzyme mapping and sequencing gDNA was extracted by homogenizing single zebrafish embryos in 50 μl of 50 mM NaOH, followed by incubation for 20 min at 95 °C, cooling to 4 °C and addition of 5 μl of 1 mM Tris-HCl pH=8 to neutralize the solution . .. The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C.

    Article Title: Genome-Wide Mapping of DNA Strand Breaks
    Article Snippet: As a negative control for DFY046 DNA end-labeling, DNA was incubated without TdT in the reaction mix and referred to as I- or N-, when using HO-induced and non-induced DNA respectively. .. After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs).

    TUNEL Assay:

    Article Title: Genome-Wide Mapping of DNA Strand Breaks
    Article Snippet: Thus, 0.5 µg of genomic DNA was incubated in TUNEL reaction mix at 37°C for 40 min in a final volume of 20 µl of TdT reaction buffer, 5 mM of CoCl2 , 0.10 mM of biotin-14-dATP (Invitrogen), 0.25 mM of dATP and 200 U of TdT. .. After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs).

    Ligation:

    Article Title: A MITE-based genotyping method to reveal hundreds of DNA polymorphisms in an animal genome after a few generations of artificial selection
    Article Snippet: .. Preparation of genomic representations For each sample, digestion and ligation reactions were carried out simultaneously at 37°C for 3 hours on 50 ng of genomic DNA, using 2 units of restriction enzyme Bsp1286I (New England Biolabs, NEB), 80 units of T4 DNA ligase (NEB) and 0.05 μM Bsp1286I adaptors I (Table ), in a buffer with final concentrations of 10 mM Tris-OAc, 50 mM KOAc, 10 mM Mg(OAc)2, 5 mM DTT (pH 7.8), 1 mM ATP and 100 ng/ml Bovine Serum Albumin (NEB). .. The obtained ligated products served as template in a first round of PCR amplification.

    Article Title: Genetic Structure, Linkage Disequilibrium and Signature of Selection in Sorghum: Lessons from Physically Anchored DArT Markers
    Article Snippet: Bsp 1286I adapters were ligated to the digested DNA with T4 DNA ligase (NEB). .. A 1-µl aliquot of the ligation product was used as the template in two amplification reactions with one DArT-Bsp 1286I primer ( 5′-GAG TAG TGC CAG AAC GGT C-3′ ) and two MITE (transposable elements) DArT-TIR specific primers ( 5′-TTT TTG GAA CTA AAC AAG GCC-3′ and 5′-G GGT GAA CTA AAC AAG GCC-3′ ).

    Generated:

    Article Title: DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis
    Article Snippet: DNA prepared by PCR was amplified with synthetic primers from synthetic templates by using Ready-To-Go PCR Beads (Amersham Biosciences), and the synthetic segments were removed from the final products by restriction digests with Bsp 1286I and Sfc I (NEB). .. PCR products and long homopolymers were generated with 5′ phosphorylation.

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: A fraction of the PCR products, including a negative control containing all reaction components except the genomic DNA, was run on a 1% agarose gel to ensure that the specific 221 base pair (bp) band was generated. .. Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele.

    Negative Control:

    Article Title: Genome-Wide Mapping of DNA Strand Breaks
    Article Snippet: As a negative control for DFY046 DNA end-labeling, DNA was incubated without TdT in the reaction mix and referred to as I- or N-, when using HO-induced and non-induced DNA respectively. .. After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs).

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: A fraction of the PCR products, including a negative control containing all reaction components except the genomic DNA, was run on a 1% agarose gel to ensure that the specific 221 base pair (bp) band was generated. .. Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele.

    Polymerase Chain Reaction:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: .. The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. ..

    Article Title: Molecular and Clinical Characterization of the Variable Phenotype in Korean Families with Hearing Loss Associated with the Mitochondrial A1555G Mutation
    Article Snippet: .. For the identification of the TRMU gene mutation G28T (A10S), the PCR amplification of exon 1 of the TRMU gene was performed using a previously reported primer, and the PCR products were analyzed using PCR-RFLP with Bsp 1286I (New England Biolabs, Ipswich, MA, USA) . ..

    Article Title: A MITE-based genotyping method to reveal hundreds of DNA polymorphisms in an animal genome after a few generations of artificial selection
    Article Snippet: Preparation of genomic representations For each sample, digestion and ligation reactions were carried out simultaneously at 37°C for 3 hours on 50 ng of genomic DNA, using 2 units of restriction enzyme Bsp1286I (New England Biolabs, NEB), 80 units of T4 DNA ligase (NEB) and 0.05 μM Bsp1286I adaptors I (Table ), in a buffer with final concentrations of 10 mM Tris-OAc, 50 mM KOAc, 10 mM Mg(OAc)2, 5 mM DTT (pH 7.8), 1 mM ATP and 100 ng/ml Bovine Serum Albumin (NEB). .. The obtained ligated products served as template in a first round of PCR amplification.

    Article Title: DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis
    Article Snippet: .. DNA prepared by PCR was amplified with synthetic primers from synthetic templates by using Ready-To-Go PCR Beads (Amersham Biosciences), and the synthetic segments were removed from the final products by restriction digests with Bsp 1286I and Sfc I (NEB). .. The resulting 70-mer (dS70 ) and 78-mer restriction products were separated by denaturing PAGE to generate single-stranded PCR products. dC500 and dA1300 were purchased from Amersham Biosciences.

    Article Title: Genetic Structure, Linkage Disequilibrium and Signature of Selection in Sorghum: Lessons from Physically Anchored DArT Markers
    Article Snippet: Bsp 1286I adapters were ligated to the digested DNA with T4 DNA ligase (NEB). .. In a first PCR, for one unit of Bsp 1286I primer, we used 10 units of TIR primers to maximize the number of different TIR fragments produced.

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR products were digested with the restriction enzyme Bsp1286 I (SduI), were separated by electrophoresis on 1,5% (w/v) agarose gel, and were visualized by ethidium bromide staining. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: .. After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. The digested products were analyzed in 1.5% agarose gel stained with ethidium bromide.

    Article Title: IMPAIRED ACTIVITY-DEPENDENT PLASTICITY OF QUANTAL AMPLITUDE AT THE NEUROMUSCULAR JUNCTION OF RAB3A DELETION AND RAB3A EARLYBIRD MUTANT MICE
    Article Snippet: .. Rab3A+/Ebd heterozygous mice were bred at Wright State University and genotyped in a two step procedure, step 1, a PCR reaction with RabF1 and Dcaps3R as primers; step 2, a digestion with enzyme Bsp1286I (New England Biolabs) that distinguishes the Earlybird mutant by its different bp products. .. TNFα−/− mice (B6.129S6-Tnftm1GKl /J) and controls (C57BL/6J) were purchased from Jackson Laboratory and housed briefly at Wright State University.

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: .. Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele. .. The restriction fragments were resolved by electrophoresis on a 3% agarose gel.

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: .. Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results. .. The SBE reaction was cycled 25 times (10sec at 96°C, 15sec at 50°C, 30sec at 60°C) and 0.05 units of shrimp alkaline phosphatase (USB Corporation, Cleveland, OH) were added to the SBE products to remove unincorporated ddNTP’s.

    Nucleic Acid Electrophoresis:

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: In RFLP analysis, the DNA sample is broken into pieces (digested) by restriction enzymes and the resulting restriction fragments are separated according to their lengths by gel electrophoresis. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results. .. Gel electrophoresis was run on an ABI Prism® 377 DNA Sequencer with the 36E-1200 Control Module and data were analyzed using GeneScan® 3.1 software (Applied Biosystems, Foster City, CA).

    In Vivo:

    Article Title: Genome-Wide Mapping of DNA Strand Breaks
    Article Snippet: Yeast DNA end-labeling To demonstrate the specific capture of DNA strand breaks induced in vivo , yeast DNA was end-labeled by TUNEL as described above with modifications. .. After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs).

    Mutagenesis:

    Article Title: Molecular and Clinical Characterization of the Variable Phenotype in Korean Families with Hearing Loss Associated with the Mitochondrial A1555G Mutation
    Article Snippet: .. For the identification of the TRMU gene mutation G28T (A10S), the PCR amplification of exon 1 of the TRMU gene was performed using a previously reported primer, and the PCR products were analyzed using PCR-RFLP with Bsp 1286I (New England Biolabs, Ipswich, MA, USA) . ..

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: .. Bsp1286I can recognize the mutation of G→C on site 263. .. Thus, PCR results showing one band of 207 bp indicate a homozygote GG with no G→C mutation.

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. Bsp1286I can recognize the mutation of G→C on site 263.

    Article Title: IMPAIRED ACTIVITY-DEPENDENT PLASTICITY OF QUANTAL AMPLITUDE AT THE NEUROMUSCULAR JUNCTION OF RAB3A DELETION AND RAB3A EARLYBIRD MUTANT MICE
    Article Snippet: .. Rab3A+/Ebd heterozygous mice were bred at Wright State University and genotyped in a two step procedure, step 1, a PCR reaction with RabF1 and Dcaps3R as primers; step 2, a digestion with enzyme Bsp1286I (New England Biolabs) that distinguishes the Earlybird mutant by its different bp products. .. TNFα−/− mice (B6.129S6-Tnftm1GKl /J) and controls (C57BL/6J) were purchased from Jackson Laboratory and housed briefly at Wright State University.

    Isolation:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. .. To identify the nature of indel mutations, PCR products were cloned into pGEM-T easy vector (Promega, #A1360) and DNA was isolated from individual clones and sequenced using an SP6 primer.

    Size-exclusion Chromatography:

    Article Title: A MITE-based genotyping method to reveal hundreds of DNA polymorphisms in an animal genome after a few generations of artificial selection
    Article Snippet: Preparation of genomic representations For each sample, digestion and ligation reactions were carried out simultaneously at 37°C for 3 hours on 50 ng of genomic DNA, using 2 units of restriction enzyme Bsp1286I (New England Biolabs, NEB), 80 units of T4 DNA ligase (NEB) and 0.05 μM Bsp1286I adaptors I (Table ), in a buffer with final concentrations of 10 mM Tris-OAc, 50 mM KOAc, 10 mM Mg(OAc)2, 5 mM DTT (pH 7.8), 1 mM ATP and 100 ng/ml Bovine Serum Albumin (NEB). .. The amplification reaction was performed with the following conditions: 94°C for 1 min; 20 cycles of 94°C for 30 sec, 50°C for 40 sec and 72°C for 1 min; followed by a final 7-min extension step at 72°C.

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR procedure was as follows: an initial denaturation step at 94°C for 2 min and then amplified for 35 cycles at 94°C for 30 sec, 57°C for 40 sec, and 72°C for 40 sec, followed by a final extension step at 72°C for 7 min. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Mouse Assay:

    Article Title: IMPAIRED ACTIVITY-DEPENDENT PLASTICITY OF QUANTAL AMPLITUDE AT THE NEUROMUSCULAR JUNCTION OF RAB3A DELETION AND RAB3A EARLYBIRD MUTANT MICE
    Article Snippet: .. Rab3A+/Ebd heterozygous mice were bred at Wright State University and genotyped in a two step procedure, step 1, a PCR reaction with RabF1 and Dcaps3R as primers; step 2, a digestion with enzyme Bsp1286I (New England Biolabs) that distinguishes the Earlybird mutant by its different bp products. .. TNFα−/− mice (B6.129S6-Tnftm1GKl /J) and controls (C57BL/6J) were purchased from Jackson Laboratory and housed briefly at Wright State University.

    Sequencing:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: Paragraph title: Restriction enzyme mapping and sequencing ... The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C.

    Article Title: Molecular and Clinical Characterization of the Variable Phenotype in Korean Families with Hearing Loss Associated with the Mitochondrial A1555G Mutation
    Article Snippet: The results were compared with the sequence of the wild-type GJB2 gene (GenBank accession number: NM_004004) to identify mutations. .. For the identification of the TRMU gene mutation G28T (A10S), the PCR amplification of exon 1 of the TRMU gene was performed using a previously reported primer, and the PCR products were analyzed using PCR-RFLP with Bsp 1286I (New England Biolabs, Ipswich, MA, USA) .

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. The judgment of gene polymorphism of D36H: The amplified gene sequence of D36H was 207 bp (from site 117 to 323) in the present study.

    Polyacrylamide Gel Electrophoresis:

    Article Title: DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis
    Article Snippet: DNA prepared by PCR was amplified with synthetic primers from synthetic templates by using Ready-To-Go PCR Beads (Amersham Biosciences), and the synthetic segments were removed from the final products by restriction digests with Bsp 1286I and Sfc I (NEB). .. The resulting 70-mer (dS70 ) and 78-mer restriction products were separated by denaturing PAGE to generate single-stranded PCR products. dC500 and dA1300 were purchased from Amersham Biosciences.

    Staining:

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR products were digested with the restriction enzyme Bsp1286 I (SduI), were separated by electrophoresis on 1,5% (w/v) agarose gel, and were visualized by ethidium bromide staining. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. The digested products were analyzed in 1.5% agarose gel stained with ethidium bromide.

    Purification:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: .. The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. ..

    Article Title: Molecular and Clinical Characterization of the Variable Phenotype in Korean Families with Hearing Loss Associated with the Mitochondrial A1555G Mutation
    Article Snippet: The PCR products were purified with the Exo-SAP enzyme (USB, Cleveland, OH, USA) and analyzed through direct sequencing. .. For the identification of the TRMU gene mutation G28T (A10S), the PCR amplification of exon 1 of the TRMU gene was performed using a previously reported primer, and the PCR products were analyzed using PCR-RFLP with Bsp 1286I (New England Biolabs, Ipswich, MA, USA) .

    Article Title: DNA heterogeneity and phosphorylation unveiled by single-molecule electrophoresis
    Article Snippet: DNA prepared by PCR was amplified with synthetic primers from synthetic templates by using Ready-To-Go PCR Beads (Amersham Biosciences), and the synthetic segments were removed from the final products by restriction digests with Bsp 1286I and Sfc I (NEB). .. All DNA except for dA1300 and dC500 were purified by PAGE under denaturing conditions.

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: Reaction C (10μl) contained 3μl of purified PCR product ( CCR5 59653) and 1.5pmol of SBE primer ( CCR5 59653). .. Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results.

    Plasmid Preparation:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. .. To identify the nature of indel mutations, PCR products were cloned into pGEM-T easy vector (Promega, #A1360) and DNA was isolated from individual clones and sequenced using an SP6 primer.

    Software:

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results. .. Gel electrophoresis was run on an ABI Prism® 377 DNA Sequencer with the 36E-1200 Control Module and data were analyzed using GeneScan® 3.1 software (Applied Biosystems, Foster City, CA).

    Multiplex Assay:

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: Each SBE reaction contained 5μl SNaPshot Multiplex Ready Reaction Mix and 1.5pmol of each SBE primer. .. Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results.

    Agarose Gel Electrophoresis:

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR products were digested with the restriction enzyme Bsp1286 I (SduI), were separated by electrophoresis on 1,5% (w/v) agarose gel, and were visualized by ethidium bromide staining. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Article Title: The Glu727 Allele of Thyroid Stimulating Hormone Receptor Gene is Associated with Osteoporosis
    Article Snippet: After amplification the PCR product was digested with fast digest restriction endonucleases, Bsp1286I and NlaIII (New England Biolabs Ltd., Pickering, ON, Canada) separately for 5 min. .. The digested products were analyzed in 1.5% agarose gel stained with ethidium bromide.

    Article Title: Shift work, hCLOCK T3111C polymorphism, and endometriosis risk
    Article Snippet: A fraction of the PCR products, including a negative control containing all reaction components except the genomic DNA, was run on a 1% agarose gel to ensure that the specific 221 base pair (bp) band was generated. .. Per the methods of Desan et al. , the PCR product was digested with Bsp1286I (New England Biolabs, Beverly, MA), which generates 95- and 126-bp bands in individuals homozygous for the C allele, 95-, 126- and 221-bp bands in heterozygous individuals, and a 221-bp band in individuals homozygous for the T allele.

    Produced:

    Article Title: Genetic Structure, Linkage Disequilibrium and Signature of Selection in Sorghum: Lessons from Physically Anchored DArT Markers
    Article Snippet: Bsp 1286I adapters were ligated to the digested DNA with T4 DNA ligase (NEB). .. In a first PCR, for one unit of Bsp 1286I primer, we used 10 units of TIR primers to maximize the number of different TIR fragments produced.

    End Labeling:

    Article Title: Genome-Wide Mapping of DNA Strand Breaks
    Article Snippet: .. After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs). ..

    CTG Assay:

    Article Title: Genetic Variant in the CYP19A1 Gene Associated with Coronary Artery Disease
    Article Snippet: The PCR amplification primers used were forward, 5′-CTG GAA CAC TAG AGA AGG CTG GTC AGT GC-3′, and reverse, 5′-GTT CTC TGG TGT GAA CAG GAG CAG ATC AC-3′. .. The Bsp1286I restriction enzyme (New England, Biolabs, Hitchin, UK) recognizes GDGCH∧ C sites (isoschizomers: SduI).

    Gel Extraction:

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering
    Article Snippet: .. The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C. ..

    Goldengate Assay:

    Article Title: Chemokine polymorphisms and lymphoma: a pooled analysis
    Article Snippet: Restriction fragment length polymorphism assays were performed on the CCR5 59029 PCR product with BSP 1286 I (New England Biolabs, Beverly, MA) on 20 random samples post-SBE genotyping to insure consistent results. .. For the NHL2 study genotyping for interim analyses[ ] was performed using the Illumina GoldenGate assay on a BeadStation 500G Genotyping System (2 SNPs in CX3 CR1 and 1 SNP in CCL5 ) as previously described.

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    New England Biolabs bsp 1286i
    miR430-mediated regulation of lft2 expression in zebrafish. ( a ) Schematic diagram of the lft2 3′UTR showing positions of TALEN-binding sites (green arrows), miR-430-binding site (MRE, red box), primers used to amplify the region (P1, P2) and the Bsp1286I restriction site (black triangle). ( b ) Detail of miR-430 MRE genomic region showing the position of TALEN-binding sites (green) and the miR-430 seed sequence (red box). ( c , d ) lft2 -miR430 TALEN-injected animals show increased lft2 expression at shield stage, as detected by in situ hybridization ( c ), or qPCR analysis of single embryos ( d ) (error bars=s.e.m. of three technical replicates). ( e ) Validation of miR-430 MRE disruption by loss of the <t>Bsp</t> <t>1286I</t> restriction site. Gel image shows PCR products of genomic DNA isolated from embryos in d digested with Bsp1286I . ( f ) Cycloptic phenotype in lft2 -miR430 TALEN-injected embryos at 50 h.p.f. (16.4% phenotypic penetrance ( n =85)). ( g ) Loss of the Bsp 1286I restriction site in genomic DNA isolated from TALEN-injected embryos in f , compared with controls. ( h ) Sequencing of region spanning the TALEN cut site from embryo in f shows discrete indels across the predicted miR-430 MRE. Seed sequence is marked by a red box, and deletions, insertions and substitutions are indicated relative to the WT sequence (first line).
    Bsp 1286i, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    miR430-mediated regulation of lft2 expression in zebrafish. ( a ) Schematic diagram of the lft2 3′UTR showing positions of TALEN-binding sites (green arrows), miR-430-binding site (MRE, red box), primers used to amplify the region (P1, P2) and the Bsp1286I restriction site (black triangle). ( b ) Detail of miR-430 MRE genomic region showing the position of TALEN-binding sites (green) and the miR-430 seed sequence (red box). ( c , d ) lft2 -miR430 TALEN-injected animals show increased lft2 expression at shield stage, as detected by in situ hybridization ( c ), or qPCR analysis of single embryos ( d ) (error bars=s.e.m. of three technical replicates). ( e ) Validation of miR-430 MRE disruption by loss of the Bsp 1286I restriction site. Gel image shows PCR products of genomic DNA isolated from embryos in d digested with Bsp1286I . ( f ) Cycloptic phenotype in lft2 -miR430 TALEN-injected embryos at 50 h.p.f. (16.4% phenotypic penetrance ( n =85)). ( g ) Loss of the Bsp 1286I restriction site in genomic DNA isolated from TALEN-injected embryos in f , compared with controls. ( h ) Sequencing of region spanning the TALEN cut site from embryo in f shows discrete indels across the predicted miR-430 MRE. Seed sequence is marked by a red box, and deletions, insertions and substitutions are indicated relative to the WT sequence (first line).

    Journal: Nature Communications

    Article Title: Understanding functional miRNA–target interactions in vivo by site-specific genome engineering

    doi: 10.1038/ncomms5640

    Figure Lengend Snippet: miR430-mediated regulation of lft2 expression in zebrafish. ( a ) Schematic diagram of the lft2 3′UTR showing positions of TALEN-binding sites (green arrows), miR-430-binding site (MRE, red box), primers used to amplify the region (P1, P2) and the Bsp1286I restriction site (black triangle). ( b ) Detail of miR-430 MRE genomic region showing the position of TALEN-binding sites (green) and the miR-430 seed sequence (red box). ( c , d ) lft2 -miR430 TALEN-injected animals show increased lft2 expression at shield stage, as detected by in situ hybridization ( c ), or qPCR analysis of single embryos ( d ) (error bars=s.e.m. of three technical replicates). ( e ) Validation of miR-430 MRE disruption by loss of the Bsp 1286I restriction site. Gel image shows PCR products of genomic DNA isolated from embryos in d digested with Bsp1286I . ( f ) Cycloptic phenotype in lft2 -miR430 TALEN-injected embryos at 50 h.p.f. (16.4% phenotypic penetrance ( n =85)). ( g ) Loss of the Bsp 1286I restriction site in genomic DNA isolated from TALEN-injected embryos in f , compared with controls. ( h ) Sequencing of region spanning the TALEN cut site from embryo in f shows discrete indels across the predicted miR-430 MRE. Seed sequence is marked by a red box, and deletions, insertions and substitutions are indicated relative to the WT sequence (first line).

    Article Snippet: The PCR product was then purified using the QIAquick Gel Extraction Kit (Qiagen, #28706) and digested with Bsp 1286I (NEB, #R0120) for 4 h at 37 °C.

    Techniques: Expressing, Binding Assay, Sequencing, Injection, In Situ Hybridization, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Isolation

    Enrichment of DNA sequences in the vicinity of three induced DSBs cleaved by the induced HO endonuclease in the yeast S. cerevisiae . ( a ) Southern blot analysis of the HO site cleavage efficiency at the recombinant PHO5-HO locus ( b ) Immunoprecipitation of DNA sequences flanking the HO cut site within PHO5-HO, digested by Bsp1286I and quantified by qPCR. ( c ) Southern blot analysis of the HO site cleavage efficiency at the mating-type locus. ( d ) Immunoprecipitation of DNA sequences flanking the HO cut site of the Mating-type locus, digested by Bsp1286I and quantified by qPCR. ( e ) Southern blot analysis of the HO site cleavage efficiency within the YcpHOcut4 locus ( f ) Immunoprecipitation of DNA sequences flanking the HO cut site of the YcpHOcut4 plasmid, digested by Bsp1286I and quantified by qPCR. Error bars represent standard deviation for 3 independent IP. I+, DNA from HO-induced cells end-labeled with dATP, biotin-dATP and TdT; I-, DNA from HO-induced cells end-labeled with dATP, biotin-dATP without TdT; N+, DNA from uninduced cells end-labeled with dATP, biotin-dATP and TdT.

    Journal: PLoS ONE

    Article Title: Genome-Wide Mapping of DNA Strand Breaks

    doi: 10.1371/journal.pone.0017353

    Figure Lengend Snippet: Enrichment of DNA sequences in the vicinity of three induced DSBs cleaved by the induced HO endonuclease in the yeast S. cerevisiae . ( a ) Southern blot analysis of the HO site cleavage efficiency at the recombinant PHO5-HO locus ( b ) Immunoprecipitation of DNA sequences flanking the HO cut site within PHO5-HO, digested by Bsp1286I and quantified by qPCR. ( c ) Southern blot analysis of the HO site cleavage efficiency at the mating-type locus. ( d ) Immunoprecipitation of DNA sequences flanking the HO cut site of the Mating-type locus, digested by Bsp1286I and quantified by qPCR. ( e ) Southern blot analysis of the HO site cleavage efficiency within the YcpHOcut4 locus ( f ) Immunoprecipitation of DNA sequences flanking the HO cut site of the YcpHOcut4 plasmid, digested by Bsp1286I and quantified by qPCR. Error bars represent standard deviation for 3 independent IP. I+, DNA from HO-induced cells end-labeled with dATP, biotin-dATP and TdT; I-, DNA from HO-induced cells end-labeled with dATP, biotin-dATP without TdT; N+, DNA from uninduced cells end-labeled with dATP, biotin-dATP and TdT.

    Article Snippet: After end-labeling, samples were precipitated as described above and dissolved in NEB4 buffer supplemented with BSA for Bsp1286I digestion (New England Biolabs).

    Techniques: Southern Blot, Recombinant, Immunoprecipitation, Real-time Polymerase Chain Reaction, Plasmid Preparation, Standard Deviation, Labeling