bsm ai  (New England Biolabs)


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    Name:
    BsmAI
    Description:
    BsmAI 5 000 units
    Catalog Number:
    r0529l
    Price:
    282
    Size:
    5 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs bsm ai
    BsmAI
    BsmAI 5 000 units
    https://www.bioz.com/result/bsm ai/product/New England Biolabs
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    bsm ai - by Bioz Stars, 2020-05
    94/100 stars

    Images

    1) Product Images from "Slow-Channel Myasthenic Syndrome Caused By Enhanced Activation, Desensitization, and Agonist Binding Affinity Attributable to Mutation in the M2 Domain of the Acetylcholine Receptor α Subunit"

    Article Title: Slow-Channel Myasthenic Syndrome Caused By Enhanced Activation, Desensitization, and Agonist Binding Affinity Attributable to Mutation in the M2 Domain of the Acetylcholine Receptor α Subunit

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.17-15-05651.1997

    Identification and analysis of mutation in the AChR α subunit. A , Automated sequencing of α exon 7 around codon 249 in a control and the propositus. In the propositus, both G and T nucleotides are present at position 745 ( arrow ), indicating a heterozygous G→T transversion. This mutation changes codon 249 from a GTC for valine to a TTC for phenylalanine. B , Allele-specific PCR and mutagenic PCR plus Bsm AI analysis of genomic DNA in the propositus’ family. Both wild-type and mutant-allele-specific primers amplify an expected 168 bp fragment in propositus and his father, but only the wild-type primer amplifies the expected fragment in the other family members. On Bsm AI analysis after mutagenic PCR, the wild-type allele gives rise to a 134 bp fragment ( open arrowhead ) and a 23 bp fragment (not shown); the mutant allele yields an undigested 157 bp fragment ( closed arrowhead ). Both wild-type and mutant fragments appear in the propositus, but only the wild-type fragment appears in other family members. The incongruity between allele-specific PCR and restriction analysis in the father is attributable to the father being a mosaic for αV249F (see text). The arrow indicates propositus. C , Multiple alignment of AChR M2 membrane-spanning domains. The boxes enclose the conserved valine residues in human AChR subunits and in AChR α subunits of other species. The mutant phenylalanine is shown at the bottom.
    Figure Legend Snippet: Identification and analysis of mutation in the AChR α subunit. A , Automated sequencing of α exon 7 around codon 249 in a control and the propositus. In the propositus, both G and T nucleotides are present at position 745 ( arrow ), indicating a heterozygous G→T transversion. This mutation changes codon 249 from a GTC for valine to a TTC for phenylalanine. B , Allele-specific PCR and mutagenic PCR plus Bsm AI analysis of genomic DNA in the propositus’ family. Both wild-type and mutant-allele-specific primers amplify an expected 168 bp fragment in propositus and his father, but only the wild-type primer amplifies the expected fragment in the other family members. On Bsm AI analysis after mutagenic PCR, the wild-type allele gives rise to a 134 bp fragment ( open arrowhead ) and a 23 bp fragment (not shown); the mutant allele yields an undigested 157 bp fragment ( closed arrowhead ). Both wild-type and mutant fragments appear in the propositus, but only the wild-type fragment appears in other family members. The incongruity between allele-specific PCR and restriction analysis in the father is attributable to the father being a mosaic for αV249F (see text). The arrow indicates propositus. C , Multiple alignment of AChR M2 membrane-spanning domains. The boxes enclose the conserved valine residues in human AChR subunits and in AChR α subunits of other species. The mutant phenylalanine is shown at the bottom.

    Techniques Used: Mutagenesis, Sequencing, Polymerase Chain Reaction

    2) Product Images from "The lss Supernodulation Mutant of Medicago truncatula Reduces Expression of the SUNN Gene 1 Gene 1 [OA]"

    Article Title: The lss Supernodulation Mutant of Medicago truncatula Reduces Expression of the SUNN Gene 1 Gene 1 [OA]

    Journal: Plant Physiology

    doi: 10.1104/pp.110.164889

    Origin of expressed SUNN in sunn LSS/ lss SUNN plants. A, Diagram of mutation and PCR product locations and restriction sites used in B. H indicates a Hae III restriction site, while B indicates a Bsm AI site; within the PCR product, differential digestion
    Figure Legend Snippet: Origin of expressed SUNN in sunn LSS/ lss SUNN plants. A, Diagram of mutation and PCR product locations and restriction sites used in B. H indicates a Hae III restriction site, while B indicates a Bsm AI site; within the PCR product, differential digestion

    Techniques Used: Mutagenesis, Polymerase Chain Reaction

    3) Product Images from "Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon"

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon

    Journal: Journal of Clinical Microbiology

    doi:

    The ITS2 region was PCR amplified with universal fungal primers ITS3 and ITS4, and the products were digested with restriction enzymes Bsm AI ( C. dubliniensis specific) (A) and Nsp BII ( C. albicans specific) (B). The restriction fragments were run on a 1.2% agarose gel and stained with GelStar (FMC Bioproducts). ∗, C. dubliniensis strains.
    Figure Legend Snippet: The ITS2 region was PCR amplified with universal fungal primers ITS3 and ITS4, and the products were digested with restriction enzymes Bsm AI ( C. dubliniensis specific) (A) and Nsp BII ( C. albicans specific) (B). The restriction fragments were run on a 1.2% agarose gel and stained with GelStar (FMC Bioproducts). ∗, C. dubliniensis strains.

    Techniques Used: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining

    Related Articles

    DNA Extraction:

    Article Title: Somatic Mosaicism in Menkes Disease Suggests Choroid Plexus-mediated Copper Transport to the Developing Brain
    Article Snippet: .. The 294 bp PCR products generated were purified using a DNA isolation kit (QIAGEN, Valencia, CA, USA), digested with BsmAI (NEB, Beverly, MA) and electrophoresed through 2% agarose. ..

    Nucleic Acid Electrophoresis:

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon
    Article Snippet: .. The PCR amplification products (∼500 ng) were digested with 1 μl of restriction enzymes Nsp BII ( C. albicans ITS2 specific) (AP Biotech, Piscataway, N.J.) and Bsm AI ( C. dubliniensis ITS2 specific) (New England Biolabs, Beverly, Mass.) for 1 h and were analyzed by gel electrophoresis in a 1.5% agarose gel. ..

    Amplification:

    Article Title: The lss Supernodulation Mutant of Medicago truncatula Reduces Expression of the SUNN Gene 1 Gene 1 [OA]
    Article Snippet: .. Following amplification, the PCR products were digested with Bsm AI (New England Biolabs) at 55°C for 4 h, which cuts only the sunn-2 allele, or Hae III (New England Biolabs) at 37°C for 4 h, which cuts only the sunn-1 allele. .. Following digestion, the samples were run on a 1% agarose gel and stained with ethidium bromide.

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon
    Article Snippet: .. The PCR amplification products (∼500 ng) were digested with 1 μl of restriction enzymes Nsp BII ( C. albicans ITS2 specific) (AP Biotech, Piscataway, N.J.) and Bsm AI ( C. dubliniensis ITS2 specific) (New England Biolabs, Beverly, Mass.) for 1 h and were analyzed by gel electrophoresis in a 1.5% agarose gel. ..

    Agarose Gel Electrophoresis:

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon
    Article Snippet: .. The PCR amplification products (∼500 ng) were digested with 1 μl of restriction enzymes Nsp BII ( C. albicans ITS2 specific) (AP Biotech, Piscataway, N.J.) and Bsm AI ( C. dubliniensis ITS2 specific) (New England Biolabs, Beverly, Mass.) for 1 h and were analyzed by gel electrophoresis in a 1.5% agarose gel. ..

    Mutagenesis:

    Article Title: High frequency of the IVS2-2A > G DNA sequence variation in SLC26A5, encoding the cochlear motor protein prestin, precludes its involvement in hereditary hearing loss
    Article Snippet: .. The presence of the 1555A > G mutation was evaluated in some subjects by DNA sequencing as previously described [ ] and in other subjects by restriction digestion using BsmAI (New England BioLabs, Beverly, MA, USA) according to manufacturer's specifications and as described [ ]. .. Sequence analysis Electropherograms were evaluated by visual inspection and pairwise alignment to reference sequences using the BCM Search Launcher BLAST2 Pairwise Sequence Alignment Tool from the Human Genome Sequencing Center of Baylor College of Medicine , and/or by interpretation using Mutation Surveyor software Version 2.41 (Softgenetics, Inc, State College, Pennsylvania, USA).

    Purification:

    Article Title: Somatic Mosaicism in Menkes Disease Suggests Choroid Plexus-mediated Copper Transport to the Developing Brain
    Article Snippet: .. The 294 bp PCR products generated were purified using a DNA isolation kit (QIAGEN, Valencia, CA, USA), digested with BsmAI (NEB, Beverly, MA) and electrophoresed through 2% agarose. ..

    Polymerase Chain Reaction:

    Article Title: Association between a Functional Polymorphism (-1195T > C) in the IGFBP5 Promoter and Head and Neck Cancer Risk
    Article Snippet: .. When the PCR products were digested with Bsma I (New England Biolabs, Beverly, MA) at 55°C overnight, the 138-bp PCR products with the -1195T allele exhibited 118- and 20-bp bands, while the C allele remained uncut; and the 120-bp PCR products with the-709C allele exhibited 98- and 22-bp bands, while the G allele remained uncut. ..

    Article Title: Slow-Channel Myasthenic Syndrome Caused By Enhanced Activation, Desensitization, and Agonist Binding Affinity Attributable to Mutation in the M2 Domain of the Acetylcholine Receptor α Subunit
    Article Snippet: .. The PCR product was digested with Bsm AI (New England Biolabs, Beverly, MA). ..

    Article Title: Somatic Mosaicism in Menkes Disease Suggests Choroid Plexus-mediated Copper Transport to the Developing Brain
    Article Snippet: .. The 294 bp PCR products generated were purified using a DNA isolation kit (QIAGEN, Valencia, CA, USA), digested with BsmAI (NEB, Beverly, MA) and electrophoresed through 2% agarose. ..

    Article Title: The lss Supernodulation Mutant of Medicago truncatula Reduces Expression of the SUNN Gene 1 Gene 1 [OA]
    Article Snippet: .. Following amplification, the PCR products were digested with Bsm AI (New England Biolabs) at 55°C for 4 h, which cuts only the sunn-2 allele, or Hae III (New England Biolabs) at 37°C for 4 h, which cuts only the sunn-1 allele. .. Following digestion, the samples were run on a 1% agarose gel and stained with ethidium bromide.

    Article Title: Pathogenicity of two COQ7 mutations and responses to 2,4‐dihydroxybenzoate bypass treatment
    Article Snippet: .. The 643‐bp PCR product was subjected to sequencing using the same primers as well as restriction fragment length polymorphism (RFLP) analysis using BsmAI (New England Biolabs). .. The digestion of PCR products from samples without the 1555A > G mutation produces two fragments of 409 and 234 bp due to the recognition of the restriction site of BsmAI.

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon
    Article Snippet: .. The PCR amplification products (∼500 ng) were digested with 1 μl of restriction enzymes Nsp BII ( C. albicans ITS2 specific) (AP Biotech, Piscataway, N.J.) and Bsm AI ( C. dubliniensis ITS2 specific) (New England Biolabs, Beverly, Mass.) for 1 h and were analyzed by gel electrophoresis in a 1.5% agarose gel. ..

    Article Title: CTLA-4 gene polymorphisms and their influence on predisposition to autoimmune thyroid diseases (Graves' disease and Hashimoto's thyroiditis)
    Article Snippet: .. Analysis of CTLA-4 polymorphism CTLA-4 polymorphisms (A49G, 1822 C/T and CT60 A/G) were assessed by PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism), using the following restriction enzymes: Fnu4HI , BsmAI , BsaAI (New England Biolabs, USA). .. First, genomic DNA sequences containing the polymorphic region, i.e., Fnu4HI or BsmAI or BsaAI restriction site, were amplified in a PCR reaction (Personal Thermocycler, Eppendorf, Germany), in a total volume of 25 µl, including: 3 µl 10x AmpliTaq Gold® 360 buffer (150 mM Tris-HCl, pH 8.3, 500 mM KCl), 0.12 µl (5 U/ µl) AmpliTaq Gold® 360 DNA Polymerase, 2 µl (25 mM) MgCl2 , 0.66 µl (10 mM) dNTPs (Applied Biosystems, USA), 1 µl (40 ng) DNA, 2.4 µl (1.2 µl 0.5 µM each primer: forward and reverse) and 15.82 µl nuclease-free water.

    Generated:

    Article Title: Somatic Mosaicism in Menkes Disease Suggests Choroid Plexus-mediated Copper Transport to the Developing Brain
    Article Snippet: .. The 294 bp PCR products generated were purified using a DNA isolation kit (QIAGEN, Valencia, CA, USA), digested with BsmAI (NEB, Beverly, MA) and electrophoresed through 2% agarose. ..

    DNA Sequencing:

    Article Title: High frequency of the IVS2-2A > G DNA sequence variation in SLC26A5, encoding the cochlear motor protein prestin, precludes its involvement in hereditary hearing loss
    Article Snippet: .. The presence of the 1555A > G mutation was evaluated in some subjects by DNA sequencing as previously described [ ] and in other subjects by restriction digestion using BsmAI (New England BioLabs, Beverly, MA, USA) according to manufacturer's specifications and as described [ ]. .. Sequence analysis Electropherograms were evaluated by visual inspection and pairwise alignment to reference sequences using the BCM Search Launcher BLAST2 Pairwise Sequence Alignment Tool from the Human Genome Sequencing Center of Baylor College of Medicine , and/or by interpretation using Mutation Surveyor software Version 2.41 (Softgenetics, Inc, State College, Pennsylvania, USA).

    Sequencing:

    Article Title: Pathogenicity of two COQ7 mutations and responses to 2,4‐dihydroxybenzoate bypass treatment
    Article Snippet: .. The 643‐bp PCR product was subjected to sequencing using the same primers as well as restriction fragment length polymorphism (RFLP) analysis using BsmAI (New England Biolabs). .. The digestion of PCR products from samples without the 1555A > G mutation produces two fragments of 409 and 234 bp due to the recognition of the restriction site of BsmAI.

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    New England Biolabs bsm ai
    Identification and analysis of mutation in the AChR α subunit. A , Automated sequencing of α exon 7 around codon 249 in a control and the propositus. In the propositus, both G and T nucleotides are present at position 745 ( arrow ), indicating a heterozygous G→T transversion. This mutation changes codon 249 from a GTC for valine to a TTC for phenylalanine. B , Allele-specific <t>PCR</t> and mutagenic PCR plus <t>Bsm</t> AI analysis of genomic DNA in the propositus’ family. Both wild-type and mutant-allele-specific primers amplify an expected 168 bp fragment in propositus and his father, but only the wild-type primer amplifies the expected fragment in the other family members. On Bsm AI analysis after mutagenic PCR, the wild-type allele gives rise to a 134 bp fragment ( open arrowhead ) and a 23 bp fragment (not shown); the mutant allele yields an undigested 157 bp fragment ( closed arrowhead ). Both wild-type and mutant fragments appear in the propositus, but only the wild-type fragment appears in other family members. The incongruity between allele-specific PCR and restriction analysis in the father is attributable to the father being a mosaic for αV249F (see text). The arrow indicates propositus. C , Multiple alignment of AChR M2 membrane-spanning domains. The boxes enclose the conserved valine residues in human AChR subunits and in AChR α subunits of other species. The mutant phenylalanine is shown at the bottom.
    Bsm Ai, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsm ai/product/New England Biolabs
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    bsm ai - by Bioz Stars, 2020-05
    94/100 stars
      Buy from Supplier

    85
    New England Biolabs restriction enzymes bsm ai
    Polymerase chain reaction (PCR)-generated double restriction enzyme sites genotyping assay. The central sequence was the reference genomic DNA sequence. This assay was a nested PCR assay where two primer sets, the outer and inner primers, were used. Each of the inner primers had one base mismatch to the reference sequence for creating a restriction site. The cutting site of <t>Bsm</t> AI (GTCTC) was created if the allele type on rs9679162 was G (corresponding to C in the other strand). The cutting site of <t>Bsp</t> MI (ACCTGC) was created if the allele type on rs6752303 was C.
    Restriction Enzymes Bsm Ai, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction enzymes bsm ai/product/New England Biolabs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    restriction enzymes bsm ai - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    Image Search Results


    Identification and analysis of mutation in the AChR α subunit. A , Automated sequencing of α exon 7 around codon 249 in a control and the propositus. In the propositus, both G and T nucleotides are present at position 745 ( arrow ), indicating a heterozygous G→T transversion. This mutation changes codon 249 from a GTC for valine to a TTC for phenylalanine. B , Allele-specific PCR and mutagenic PCR plus Bsm AI analysis of genomic DNA in the propositus’ family. Both wild-type and mutant-allele-specific primers amplify an expected 168 bp fragment in propositus and his father, but only the wild-type primer amplifies the expected fragment in the other family members. On Bsm AI analysis after mutagenic PCR, the wild-type allele gives rise to a 134 bp fragment ( open arrowhead ) and a 23 bp fragment (not shown); the mutant allele yields an undigested 157 bp fragment ( closed arrowhead ). Both wild-type and mutant fragments appear in the propositus, but only the wild-type fragment appears in other family members. The incongruity between allele-specific PCR and restriction analysis in the father is attributable to the father being a mosaic for αV249F (see text). The arrow indicates propositus. C , Multiple alignment of AChR M2 membrane-spanning domains. The boxes enclose the conserved valine residues in human AChR subunits and in AChR α subunits of other species. The mutant phenylalanine is shown at the bottom.

    Journal: The Journal of Neuroscience

    Article Title: Slow-Channel Myasthenic Syndrome Caused By Enhanced Activation, Desensitization, and Agonist Binding Affinity Attributable to Mutation in the M2 Domain of the Acetylcholine Receptor α Subunit

    doi: 10.1523/JNEUROSCI.17-15-05651.1997

    Figure Lengend Snippet: Identification and analysis of mutation in the AChR α subunit. A , Automated sequencing of α exon 7 around codon 249 in a control and the propositus. In the propositus, both G and T nucleotides are present at position 745 ( arrow ), indicating a heterozygous G→T transversion. This mutation changes codon 249 from a GTC for valine to a TTC for phenylalanine. B , Allele-specific PCR and mutagenic PCR plus Bsm AI analysis of genomic DNA in the propositus’ family. Both wild-type and mutant-allele-specific primers amplify an expected 168 bp fragment in propositus and his father, but only the wild-type primer amplifies the expected fragment in the other family members. On Bsm AI analysis after mutagenic PCR, the wild-type allele gives rise to a 134 bp fragment ( open arrowhead ) and a 23 bp fragment (not shown); the mutant allele yields an undigested 157 bp fragment ( closed arrowhead ). Both wild-type and mutant fragments appear in the propositus, but only the wild-type fragment appears in other family members. The incongruity between allele-specific PCR and restriction analysis in the father is attributable to the father being a mosaic for αV249F (see text). The arrow indicates propositus. C , Multiple alignment of AChR M2 membrane-spanning domains. The boxes enclose the conserved valine residues in human AChR subunits and in AChR α subunits of other species. The mutant phenylalanine is shown at the bottom.

    Article Snippet: The PCR product was digested with Bsm AI (New England Biolabs, Beverly, MA).

    Techniques: Mutagenesis, Sequencing, Polymerase Chain Reaction

    Origin of expressed SUNN in sunn LSS/ lss SUNN plants. A, Diagram of mutation and PCR product locations and restriction sites used in B. H indicates a Hae III restriction site, while B indicates a Bsm AI site; within the PCR product, differential digestion

    Journal: Plant Physiology

    Article Title: The lss Supernodulation Mutant of Medicago truncatula Reduces Expression of the SUNN Gene 1 Gene 1 [OA]

    doi: 10.1104/pp.110.164889

    Figure Lengend Snippet: Origin of expressed SUNN in sunn LSS/ lss SUNN plants. A, Diagram of mutation and PCR product locations and restriction sites used in B. H indicates a Hae III restriction site, while B indicates a Bsm AI site; within the PCR product, differential digestion

    Article Snippet: Following amplification, the PCR products were digested with Bsm AI (New England Biolabs) at 55°C for 4 h, which cuts only the sunn-2 allele, or Hae III (New England Biolabs) at 37°C for 4 h, which cuts only the sunn-1 allele.

    Techniques: Mutagenesis, Polymerase Chain Reaction

    The ITS2 region was PCR amplified with universal fungal primers ITS3 and ITS4, and the products were digested with restriction enzymes Bsm AI ( C. dubliniensis specific) (A) and Nsp BII ( C. albicans specific) (B). The restriction fragments were run on a 1.2% agarose gel and stained with GelStar (FMC Bioproducts). ∗, C. dubliniensis strains.

    Journal: Journal of Clinical Microbiology

    Article Title: Rapid Identification of Candida dubliniensis Using a Species-Specific Molecular Beacon

    doi:

    Figure Lengend Snippet: The ITS2 region was PCR amplified with universal fungal primers ITS3 and ITS4, and the products were digested with restriction enzymes Bsm AI ( C. dubliniensis specific) (A) and Nsp BII ( C. albicans specific) (B). The restriction fragments were run on a 1.2% agarose gel and stained with GelStar (FMC Bioproducts). ∗, C. dubliniensis strains.

    Article Snippet: The PCR amplification products (∼500 ng) were digested with 1 μl of restriction enzymes Nsp BII ( C. albicans ITS2 specific) (AP Biotech, Piscataway, N.J.) and Bsm AI ( C. dubliniensis ITS2 specific) (New England Biolabs, Beverly, Mass.) for 1 h and were analyzed by gel electrophoresis in a 1.5% agarose gel.

    Techniques: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Staining

    Polymerase chain reaction (PCR)-generated double restriction enzyme sites genotyping assay. The central sequence was the reference genomic DNA sequence. This assay was a nested PCR assay where two primer sets, the outer and inner primers, were used. Each of the inner primers had one base mismatch to the reference sequence for creating a restriction site. The cutting site of Bsm AI (GTCTC) was created if the allele type on rs9679162 was G (corresponding to C in the other strand). The cutting site of Bsp MI (ACCTGC) was created if the allele type on rs6752303 was C.

    Journal: Oncology Letters

    Article Title: Genotyping the GALNT14 gene by joint analysis of two linked single nucleotide polymorphisms using liver tissues for clinical and geographical comparisons

    doi: 10.3892/ol.2014.2507

    Figure Lengend Snippet: Polymerase chain reaction (PCR)-generated double restriction enzyme sites genotyping assay. The central sequence was the reference genomic DNA sequence. This assay was a nested PCR assay where two primer sets, the outer and inner primers, were used. Each of the inner primers had one base mismatch to the reference sequence for creating a restriction site. The cutting site of Bsm AI (GTCTC) was created if the allele type on rs9679162 was G (corresponding to C in the other strand). The cutting site of Bsp MI (ACCTGC) was created if the allele type on rs6752303 was C.

    Article Snippet: The reaction mixture comprised the second amplicon (3 μl), 10X buffer (2 μl), restriction enzymes Bsm AI or Bsp MI (1 μl) (New England Biolab, Ipswich, MA, USA) and water (14 μl).

    Techniques: Polymerase Chain Reaction, Generated, Genotyping Assay, Sequencing, Nested PCR