bst ui  (New England Biolabs)


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    Name:
    BstUI
    Description:
    BstUI 5 000 units
    Catalog Number:
    r0518l
    Price:
    257
    Size:
    5 000 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs bst ui
    BstUI
    BstUI 5 000 units
    https://www.bioz.com/result/bst ui/product/New England Biolabs
    Average 99 stars, based on 56 article reviews
    Price from $9.99 to $1999.99
    bst ui - by Bioz Stars, 2020-08
    99/100 stars

    Images

    1) Product Images from "Human Placental-Specific Epipolymorphism and its Association with Adverse Pregnancy Outcomes"

    Article Title: Human Placental-Specific Epipolymorphism and its Association with Adverse Pregnancy Outcomes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0007389

    Promoter CpG methylation correlates with lack of TUSC3 gene expression. (A) Schematic of TUSC3 locus showing the regions investigated by bisulfite pyrosequencing on the promoter (−105 to −57 relative to the transcriptional start site according to NM_006765) and genotyping assays of the 5′ untranslated region. PCR primers for DNA genotyping are indicated by black arrows while RT-PCR primers for mRNA genotyping are indicated by arrows highlighted in white. Enzyme recognition sites for Bst UI are indicated by “ B ”. (B) Methylation status of TUSC3 promoter region studied by bisulfite pyrosequencing. A similar methylation level of every CpG within each sample is observed and the gene follows “on-or-off” methylation pattern. Each circle represents a CpG site in a sample. Area shaded in black is proportional to the methylation level of the CpG site indicated by pyrosequencing. (C) Validation of complete methylation-sensitive restriction enzyme digestion on unmethylated molecules. Genomic DNA was predigested with Bst UI followed by PCR amplification with TUSC3 and ID2 specific primers ( Table S2 ). Bst UI digestion sites within the ID2 region were unmethylated ( Figure S6 ) and, therefore, no PCR product was generated after enzyme digestion. (D) Allele-specific methylation of TUSC3 on the fragment containing SNP rs12550009 demonstrated by enzyme digestion pyrosequencing. The “Simplex” diagrams (top) show the reference pyrograms by genotype. A heterozygous CT in the methylated samples (PM55 and PM123) displays a homozygous T pattern after Bst UI digestion indicating predominant methylation of the T allele. Allele-specific mRNA expression is concordant with allele-specific methylation on the same SNP rs1250009. Predominant expression of C alleles was observed in the cDNAs generated by RNA specific primers (bottom). RT+ and RT− represent assays with Reverse Transcriptase and without Reverse Transcriptase, respectively.
    Figure Legend Snippet: Promoter CpG methylation correlates with lack of TUSC3 gene expression. (A) Schematic of TUSC3 locus showing the regions investigated by bisulfite pyrosequencing on the promoter (−105 to −57 relative to the transcriptional start site according to NM_006765) and genotyping assays of the 5′ untranslated region. PCR primers for DNA genotyping are indicated by black arrows while RT-PCR primers for mRNA genotyping are indicated by arrows highlighted in white. Enzyme recognition sites for Bst UI are indicated by “ B ”. (B) Methylation status of TUSC3 promoter region studied by bisulfite pyrosequencing. A similar methylation level of every CpG within each sample is observed and the gene follows “on-or-off” methylation pattern. Each circle represents a CpG site in a sample. Area shaded in black is proportional to the methylation level of the CpG site indicated by pyrosequencing. (C) Validation of complete methylation-sensitive restriction enzyme digestion on unmethylated molecules. Genomic DNA was predigested with Bst UI followed by PCR amplification with TUSC3 and ID2 specific primers ( Table S2 ). Bst UI digestion sites within the ID2 region were unmethylated ( Figure S6 ) and, therefore, no PCR product was generated after enzyme digestion. (D) Allele-specific methylation of TUSC3 on the fragment containing SNP rs12550009 demonstrated by enzyme digestion pyrosequencing. The “Simplex” diagrams (top) show the reference pyrograms by genotype. A heterozygous CT in the methylated samples (PM55 and PM123) displays a homozygous T pattern after Bst UI digestion indicating predominant methylation of the T allele. Allele-specific mRNA expression is concordant with allele-specific methylation on the same SNP rs1250009. Predominant expression of C alleles was observed in the cDNAs generated by RNA specific primers (bottom). RT+ and RT− represent assays with Reverse Transcriptase and without Reverse Transcriptase, respectively.

    Techniques Used: CpG Methylation Assay, Expressing, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Methylation, Amplification, Generated

    2) Product Images from "Human Topoisomerase I Promotes Initiation of Simian Virus 40 DNA Replication In Vitro"

    Article Title: Human Topoisomerase I Promotes Initiation of Simian Virus 40 DNA Replication In Vitro

    Journal: Molecular and Cellular Biology

    doi:

    Competitive in vitro DNA replication reactions. (A) Competition with 1-246T. For lanes 1 to 3, 1 μg of immunoaffinity-purified T antigen was used to program an in vitro DNA replication reaction of 236 ng of pSV0II+ and 1 μCi of [α- 32 P]dATP in the absence (lanes 1 and 2) or presence (lane 3) of 100 ng of human topo I; lane 1 is a negative control without T antigen. For lanes 4 to 7, T-antigen-mediated in vitro DNA replication was initiated without label for 30 min. Then 1 μCi of [α- 32 P]dATP, 100 ng of human topo I (lanes 5 and 6), and a 10 M excess (over WT T antigen) of 1-246T (lanes 6 and 7) were added, and the reaction continued for an additional 30 min. The labeled DNA was purified, analyzed on a 1.5% agarose gel, dried, and exposed to film. The radioactive products were excised and subjected to Cerenkov counting. (B) Stimulation of completed molecules and overall DNA replication in the presence of topo I (−1-246T) or in the presence of topo I and 1-246T (+1-246T) DNA replication in the presence of T antigen alone is given a value of 1, and fold stimulation is calculated for each molar excess of 1-246T over WT T antigen and depicted graphically. The experiment containing 0.6 M equivalents of 1-246T (relative to WT T antigen) was done with a delayed pulse-chase assay. Experiments containing 2 and 10 M excesses of 1-246T were performed with delayed pulse assays; 10 M excess represents the experiment illustrated in panel A. (C) Restriction enzyme digestion of the form I molecules obtained during a delayed pulse competition reaction. A competition reaction with a 10 M excess of 1-246T was carried out as for panel A. After separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. The 739-bp fragment contains the origin of replication (ori), and the 493- and 330-bp fragments are on the opposite side close to the site of termination. The numbers below the lanes represent relative incorporation into form I DNA. (D) Competition with the catalytic site topo I mutant Y723F. An in vitro DNA replication assay was performed in the presence or absence of various quantities of WT human topo I and mutant topo I Y723F. The DNA was synthesized in the presence of label for 1 h and analyzed on a 1.5% agarose gel. Replicated completed molecules (from CCC to form I on the gel) were quantitated, and the results (from two independent experiments) are expressed as a percentage of the values obtained with WT topo I alone. Samples: 1, no T antigen (negative control); 2, T antigen alone; 3, 100 ng of human topo I; 4, 50 ng of topo I and 50 ng of Y723F; 5, 150 ng of topo I; 6, 50 ng of topo I and 100 ng of Y723F. Samples 1, 2, and 4 were normalized to sample 3, and sample 6 was normalized to sample 5.
    Figure Legend Snippet: Competitive in vitro DNA replication reactions. (A) Competition with 1-246T. For lanes 1 to 3, 1 μg of immunoaffinity-purified T antigen was used to program an in vitro DNA replication reaction of 236 ng of pSV0II+ and 1 μCi of [α- 32 P]dATP in the absence (lanes 1 and 2) or presence (lane 3) of 100 ng of human topo I; lane 1 is a negative control without T antigen. For lanes 4 to 7, T-antigen-mediated in vitro DNA replication was initiated without label for 30 min. Then 1 μCi of [α- 32 P]dATP, 100 ng of human topo I (lanes 5 and 6), and a 10 M excess (over WT T antigen) of 1-246T (lanes 6 and 7) were added, and the reaction continued for an additional 30 min. The labeled DNA was purified, analyzed on a 1.5% agarose gel, dried, and exposed to film. The radioactive products were excised and subjected to Cerenkov counting. (B) Stimulation of completed molecules and overall DNA replication in the presence of topo I (−1-246T) or in the presence of topo I and 1-246T (+1-246T) DNA replication in the presence of T antigen alone is given a value of 1, and fold stimulation is calculated for each molar excess of 1-246T over WT T antigen and depicted graphically. The experiment containing 0.6 M equivalents of 1-246T (relative to WT T antigen) was done with a delayed pulse-chase assay. Experiments containing 2 and 10 M excesses of 1-246T were performed with delayed pulse assays; 10 M excess represents the experiment illustrated in panel A. (C) Restriction enzyme digestion of the form I molecules obtained during a delayed pulse competition reaction. A competition reaction with a 10 M excess of 1-246T was carried out as for panel A. After separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. The 739-bp fragment contains the origin of replication (ori), and the 493- and 330-bp fragments are on the opposite side close to the site of termination. The numbers below the lanes represent relative incorporation into form I DNA. (D) Competition with the catalytic site topo I mutant Y723F. An in vitro DNA replication assay was performed in the presence or absence of various quantities of WT human topo I and mutant topo I Y723F. The DNA was synthesized in the presence of label for 1 h and analyzed on a 1.5% agarose gel. Replicated completed molecules (from CCC to form I on the gel) were quantitated, and the results (from two independent experiments) are expressed as a percentage of the values obtained with WT topo I alone. Samples: 1, no T antigen (negative control); 2, T antigen alone; 3, 100 ng of human topo I; 4, 50 ng of topo I and 50 ng of Y723F; 5, 150 ng of topo I; 6, 50 ng of topo I and 100 ng of Y723F. Samples 1, 2, and 4 were normalized to sample 3, and sample 6 was normalized to sample 5.

    Techniques Used: In Vitro, Purification, Negative Control, Labeling, Agarose Gel Electrophoresis, Pulse Chase, Mutagenesis, Synthesized, Countercurrent Chromatography

    3) Product Images from "5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation"

    Article Title: 5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0103714

    Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).
    Figure Legend Snippet: Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).

    Techniques Used: Methylation, Generated, Polymerase Chain Reaction, Fluorescence

    4) Product Images from "Epigenetic Silencing of the Protocadherin Family Member PCDH-γ-A11 in Astrocytomas 1"

    Article Title: Epigenetic Silencing of the Protocadherin Family Member PCDH-γ-A11 in Astrocytomas 1

    Journal: Neoplasia (New York, N.Y.)

    doi:

    Methylation analysis of a CpG island within the first exon of the PCDH-γ-A11 gene. (A) Bisulfite sequencing profile of a WHO grade II astrocytoma (7282) and normal brain (grey matter 12434). Methylated CpG sites 6 to 14 are indicated by arrows. Underlined are three Bst UI restriction sites that are methylated and therefore conserved during bisulfite treatment in this tumor sample and are used for the restriction-based methylation assay. (B) Bisulfite sequencing analysis of CpG sites 1 to 26 of individual clones of the 312-bp PCDH-γ-A11 PCR product of four tumor samples (AII 7282, AII 11092, AII 12020, and GBM 4732) and two normal brain samples (white matter 12768, grey matter 12434). (▪) Methylated CpG site; (□) unmethylated CpG site. (C) Restriction-based methylation assay of the first exon of PCDH -γ- A11 in astrocytomas. The 312-bp bisulfite PCR products without (-) and after treatment (+) with Bst UI restriction enzyme are shown. Restricted fragments of the PCR product are only apparent in tumor samples and not in the normal brain tissues (NB1, white matter 12284; NB2, white matter 12692; NB3, white matter 12768). Tumor samples AIII 2446, AIII 10414, and GBM 11637 also show no methylation in this assay.
    Figure Legend Snippet: Methylation analysis of a CpG island within the first exon of the PCDH-γ-A11 gene. (A) Bisulfite sequencing profile of a WHO grade II astrocytoma (7282) and normal brain (grey matter 12434). Methylated CpG sites 6 to 14 are indicated by arrows. Underlined are three Bst UI restriction sites that are methylated and therefore conserved during bisulfite treatment in this tumor sample and are used for the restriction-based methylation assay. (B) Bisulfite sequencing analysis of CpG sites 1 to 26 of individual clones of the 312-bp PCDH-γ-A11 PCR product of four tumor samples (AII 7282, AII 11092, AII 12020, and GBM 4732) and two normal brain samples (white matter 12768, grey matter 12434). (▪) Methylated CpG site; (□) unmethylated CpG site. (C) Restriction-based methylation assay of the first exon of PCDH -γ- A11 in astrocytomas. The 312-bp bisulfite PCR products without (-) and after treatment (+) with Bst UI restriction enzyme are shown. Restricted fragments of the PCR product are only apparent in tumor samples and not in the normal brain tissues (NB1, white matter 12284; NB2, white matter 12692; NB3, white matter 12768). Tumor samples AIII 2446, AIII 10414, and GBM 11637 also show no methylation in this assay.

    Techniques Used: Methylation, Methylation Sequencing, Clone Assay, Polymerase Chain Reaction

    5) Product Images from "SIP1 is downregulated in hepatocellular carcinoma by promoter hypermethylation"

    Article Title: SIP1 is downregulated in hepatocellular carcinoma by promoter hypermethylation

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-11-223

    Methylation analysis of promoter regions by COBRA . Photographs are representative of tumor-specific methylation in three promoter regions. Amplicons of P1 and P2 are cut with Bst UI (left and middle), and Taq I digestion is applied to the PCR products of the P3 region (right). N: normal; T: tumor; M: marker.
    Figure Legend Snippet: Methylation analysis of promoter regions by COBRA . Photographs are representative of tumor-specific methylation in three promoter regions. Amplicons of P1 and P2 are cut with Bst UI (left and middle), and Taq I digestion is applied to the PCR products of the P3 region (right). N: normal; T: tumor; M: marker.

    Techniques Used: Methylation, Combined Bisulfite Restriction Analysis Assay, Polymerase Chain Reaction, Marker

    6) Product Images from "Population Structure of Alexandrium (Dinophyceae) Cyst Formation-Promoting Bacteria in Hiroshima Bay, Japan"

    Article Title: Population Structure of Alexandrium (Dinophyceae) Cyst Formation-Promoting Bacteria in Hiroshima Bay, Japan

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.69.11.6560-6568.2003

    Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with Rsa I (B), Bst UI (C), Mbo I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.
    Figure Legend Snippet: Amplified products of the 16S rDNA of various Alex-CFPB (A) and digestion of the PCR products with Rsa I (B), Bst UI (C), Mbo I (D), and Hha I (E). Lane M, 100-bp ladder markers; lane 1, CFPB-A9; lane 2, CFPB-B1; lane 3, CFPB-C1; lane 4, CFPB-D1; lane 5, CFPB-E1; lane 6, CFPB-F1; lane 7, CFPB-G1; lane 8, CFPB-H1; lane 9, CFPB-I1; lane 10, CFPB-J1; lane 11, CFPB-K1; lane 12, CFPB-L1; lane 13, CFPB-M1; lane 14, CFPB-N1.

    Techniques Used: Amplification, Polymerase Chain Reaction

    7) Product Images from "Simultaneous detection of CpG methylation and single nucleotide polymorphism by denaturing high performance liquid chromatography"

    Article Title: Simultaneous detection of CpG methylation and single nucleotide polymorphism by denaturing high performance liquid chromatography

    Journal: Nucleic Acids Research

    doi:

    Detection of methylation in the hMLH1 promoter by Bst UI COBRA assay in the RKO and PACM82 cell lines. DNA without or with bisulfite treatment was amplified by PCR or ssPCR. The amplicon mixture was digested with Bst UI at 60°C for 3 h. M1, PCR and ssPCR as in Figure 1.
    Figure Legend Snippet: Detection of methylation in the hMLH1 promoter by Bst UI COBRA assay in the RKO and PACM82 cell lines. DNA without or with bisulfite treatment was amplified by PCR or ssPCR. The amplicon mixture was digested with Bst UI at 60°C for 3 h. M1, PCR and ssPCR as in Figure 1.

    Techniques Used: Methylation, Combined Bisulfite Restriction Analysis Assay, Amplification, Polymerase Chain Reaction

    8) Product Images from "5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation"

    Article Title: 5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0103714

    Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).
    Figure Legend Snippet: Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).

    Techniques Used: Methylation, Generated, Polymerase Chain Reaction, Fluorescence

    9) Product Images from "c-Myc Acts as a Competing Endogenous RNA to Sponge miR-34a, in the Upregulation of CD44, in Urothelial Carcinoma"

    Article Title: c-Myc Acts as a Competing Endogenous RNA to Sponge miR-34a, in the Upregulation of CD44, in Urothelial Carcinoma

    Journal: Cancers

    doi: 10.3390/cancers11101457

    miR-34a is epigenetically silenced by promoter DNA methylation in UMUC3 cell lines, and a subset of urothelial carcinoma (UC) patient samples. ( A ) Expression of pri- miR-34a in normal HUC1 bladder cells and various UC cell lines, relative to GAPDH, as an internal control. Methylation analysis of the miR-34a promoter in various UC cell lines, using ( B ) bisulphite pyrosequencing and ( C ) COBRA assay. The upper panel in ( B ) presents the genomic structure of the miR-34a promoter, with the corresponding locations of all CpG sites from −200 to +400. The lower panel in ( B ) illustrates DNA methylation at each CpG site (circle) of various cells, where the intensity of the blue color indicates the degree of methylation. The CpG sites interrogated by bisulphite prosequencing are also indicated. In the COBRA assays in ( C ), bisulphite-modified DNA was amplified via PCR and digested using Bst UI. U, undigested control; C, digested using Bst UI; M, DNA ladder marker; IVD , in vitro methylated DNA. ( D ) Expression of pri -miR-34a in UC cell lines, following epigenetic treatment. UC cell lines treated with 5-aza-2’-deoxycytidine (5aza) and/or trichostatin A (TSA) were examined for pri -miR-34a expression, using qRT-PCR. Error bars represent standard deviations calculated from duplicates. ( E ) Bisulphite pyrosequencing analysis of the miR-34a promoter in UMUC3 cells treated with various epigenetic drugs, as in ( D ). ( F ) Scatter plot showing expression and promoter methylation of miR-34a in 55 urothelial carcinoma patient samples, showing a distinct group of patient samples (red dots) with both low expression and low promoter methylation of miR-34a . Analysis of the other patient samples (methylated) showed an inverse correlation (r = −0.31) between expression and methylation of miR-34a , as shown in ( G ).
    Figure Legend Snippet: miR-34a is epigenetically silenced by promoter DNA methylation in UMUC3 cell lines, and a subset of urothelial carcinoma (UC) patient samples. ( A ) Expression of pri- miR-34a in normal HUC1 bladder cells and various UC cell lines, relative to GAPDH, as an internal control. Methylation analysis of the miR-34a promoter in various UC cell lines, using ( B ) bisulphite pyrosequencing and ( C ) COBRA assay. The upper panel in ( B ) presents the genomic structure of the miR-34a promoter, with the corresponding locations of all CpG sites from −200 to +400. The lower panel in ( B ) illustrates DNA methylation at each CpG site (circle) of various cells, where the intensity of the blue color indicates the degree of methylation. The CpG sites interrogated by bisulphite prosequencing are also indicated. In the COBRA assays in ( C ), bisulphite-modified DNA was amplified via PCR and digested using Bst UI. U, undigested control; C, digested using Bst UI; M, DNA ladder marker; IVD , in vitro methylated DNA. ( D ) Expression of pri -miR-34a in UC cell lines, following epigenetic treatment. UC cell lines treated with 5-aza-2’-deoxycytidine (5aza) and/or trichostatin A (TSA) were examined for pri -miR-34a expression, using qRT-PCR. Error bars represent standard deviations calculated from duplicates. ( E ) Bisulphite pyrosequencing analysis of the miR-34a promoter in UMUC3 cells treated with various epigenetic drugs, as in ( D ). ( F ) Scatter plot showing expression and promoter methylation of miR-34a in 55 urothelial carcinoma patient samples, showing a distinct group of patient samples (red dots) with both low expression and low promoter methylation of miR-34a . Analysis of the other patient samples (methylated) showed an inverse correlation (r = −0.31) between expression and methylation of miR-34a , as shown in ( G ).

    Techniques Used: DNA Methylation Assay, Expressing, Methylation, Combined Bisulfite Restriction Analysis Assay, Modification, Amplification, Polymerase Chain Reaction, Marker, In Vitro, Quantitative RT-PCR

    10) Product Images from "Clade-Specific 16S Ribosomal DNA Oligonucleotides Reveal the Predominance of a Single Marine Synechococcus Clade throughout a Stratified Water Column in the Red Sea"

    Article Title: Clade-Specific 16S Ribosomal DNA Oligonucleotides Reveal the Predominance of a Single Marine Synechococcus Clade throughout a Stratified Water Column in the Red Sea

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.69.5.2430-2443.2003

    RFLP analysis of Synechococcus sp. isolates from the Red Sea. PCR amplicons of ntcA were digested with Hae III and Bst UI and separated by electrophoresis on a 10% polyacrylamide gel, revealing six different RFLP patterns. Patterns from other Synechococcus strains are shown for comparison. Lanes 1 to 14: Synechococcus sp. strains RS9901, RS9902, RS9903, RS9904, RS9905, RS9907, RS9908, RS9910, RS9911, RS9915, RS9916, RS9919, RS9920, and RS9921, respectively. Lanes 15 to 19: Synechococcus sp. strains WH 7803, WH 8018, WH 8103, Minos01, and CC9311, respectively.
    Figure Legend Snippet: RFLP analysis of Synechococcus sp. isolates from the Red Sea. PCR amplicons of ntcA were digested with Hae III and Bst UI and separated by electrophoresis on a 10% polyacrylamide gel, revealing six different RFLP patterns. Patterns from other Synechococcus strains are shown for comparison. Lanes 1 to 14: Synechococcus sp. strains RS9901, RS9902, RS9903, RS9904, RS9905, RS9907, RS9908, RS9910, RS9911, RS9915, RS9916, RS9919, RS9920, and RS9921, respectively. Lanes 15 to 19: Synechococcus sp. strains WH 7803, WH 8018, WH 8103, Minos01, and CC9311, respectively.

    Techniques Used: Polymerase Chain Reaction, Electrophoresis

    Related Articles

    Agarose Gel Electrophoresis:

    Article Title: Human Topoisomerase I Promotes Initiation of Simian Virus 40 DNA Replication In Vitro
    Article Snippet: .. In some cases, after separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. .. Pulse-chase assays were performed by a modification of the procedures described by Fotedar et al. ( ) (see Fig. A).

    In Vitro:

    Article Title: High-Risk Human Papillomavirus E7 Alters Host DNA Methylome and Represses HLA-E Expression in Human Keratinocytes
    Article Snippet: .. Control DNA was generated by in vitro DNA methylation of gDNA extracted from W12E, W12G, and W12GPXY cells using McrBC endonuclease or M.SssI CpG methyltransferase followed by BstUI digestion (New England Biolabs). .. In vitro DNA methylation was performed using the M.SssI CpG methyltransferase and methylation efficiency was validated by McrBC or BstUI digestion.

    Mutagenesis:

    Article Title: Recurrent mutations in a SERPINC1 hotspot associate with venous thrombosis without apparent antithrombin deficiency
    Article Snippet: .. Identification of the c.883G > A mutation was done by restriction analysis of the 649-bp PCR product with BstUI (New England Biolabs, Ipswich, MA, USA). .. Briefly, 10 μL PCR product was completely digested with 2 U of BstUI during 2 hours at 60°C.

    Incubation:

    Article Title: Evidence for Transgenerational Transmission of Epigenetic Tumor Susceptibility in Drosophila
    Article Snippet: .. For restriction digests, 3 μg of genomic DNA was incubated with 10 units of BstUI (New England Biolabs, http://www.neb.com/ ) or 10 units of HaeIII (New England Biolabs) in 60 μl of total volume at 37 °C. ..

    Purification:

    Article Title: Human Topoisomerase I Promotes Initiation of Simian Virus 40 DNA Replication In Vitro
    Article Snippet: .. In some cases, after separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. .. Pulse-chase assays were performed by a modification of the procedures described by Fotedar et al. ( ) (see Fig. A).

    Generated:

    Article Title: High-Risk Human Papillomavirus E7 Alters Host DNA Methylome and Represses HLA-E Expression in Human Keratinocytes
    Article Snippet: .. Control DNA was generated by in vitro DNA methylation of gDNA extracted from W12E, W12G, and W12GPXY cells using McrBC endonuclease or M.SssI CpG methyltransferase followed by BstUI digestion (New England Biolabs). .. In vitro DNA methylation was performed using the M.SssI CpG methyltransferase and methylation efficiency was validated by McrBC or BstUI digestion.

    Polymerase Chain Reaction:

    Article Title: Choline availability modulates human neuroblastoma cell proliferation and alters the methylation of the promoter region of the cyclin-dependent kinase inhibitor 3 gene
    Article Snippet: .. Aliquots (30 μL) of the PCR-amplified products were digested with restriction enzymes, 40 units of BstU I at 60°C for 4 h (5′…cg^cg…3′; New England Biolabs, Beverly, MA, USA). .. Then, the products were subjected to electrophoresis on a 3% agarose gel and visualized by SYBR Green I (BMA).

    Article Title: Homozygosity for Pro of p53 Arg72Pro as a potential risk factor for hepatocellular carcinoma in Chinese population
    Article Snippet: .. PCR condition was 2 min at 94 °C, followed by 35 cycles of 30 s at 94 °C, 30 s at 60 °C, and 30 s at 72 °C, and with a final extension at 72 °C for 7 min. A 10-μL aliquot of PCR product was digested overnight at 60 °C in a 15-μL reaction volume containing 10 units of BstU I (New England BioLabs). .. After overnight digestion, the fragments were separated by electrophoresis on a vertical 90 g/L non-denaturing polyacrylamide gel at 120 V for 45 min, stained with ethidium bromide.

    Article Title: Recurrent mutations in a SERPINC1 hotspot associate with venous thrombosis without apparent antithrombin deficiency
    Article Snippet: .. Identification of the c.883G > A mutation was done by restriction analysis of the 649-bp PCR product with BstUI (New England Biolabs, Ipswich, MA, USA). .. Briefly, 10 μL PCR product was completely digested with 2 U of BstUI during 2 hours at 60°C.

    Article Title: Lack of HLA class II antigen expression in microsatellite unstable colorectal carcinomas is caused by mutations in HLA class II regulatory genes
    Article Snippet: .. The PCR products were digested with BstUI (New England Biolabs, Frankfurt, Germany). .. DNA from the cell line HCT116 was included as a positive control for CIITA promoter IV methylation in each assay.

    DNA Methylation Assay:

    Article Title: High-Risk Human Papillomavirus E7 Alters Host DNA Methylome and Represses HLA-E Expression in Human Keratinocytes
    Article Snippet: .. Control DNA was generated by in vitro DNA methylation of gDNA extracted from W12E, W12G, and W12GPXY cells using McrBC endonuclease or M.SssI CpG methyltransferase followed by BstUI digestion (New England Biolabs). .. In vitro DNA methylation was performed using the M.SssI CpG methyltransferase and methylation efficiency was validated by McrBC or BstUI digestion.

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    New England Biolabs bst ui
    Promoter CpG methylation correlates with lack of TUSC3 gene expression. (A) Schematic of TUSC3 locus showing the regions investigated by bisulfite pyrosequencing on the promoter (−105 to −57 relative to the transcriptional start site according to NM_006765) and genotyping assays of the 5′ untranslated region. PCR primers for <t>DNA</t> genotyping are indicated by black arrows while RT-PCR primers for mRNA genotyping are indicated by arrows highlighted in white. Enzyme recognition sites for <t>Bst</t> UI are indicated by “ B ”. (B) Methylation status of TUSC3 promoter region studied by bisulfite pyrosequencing. A similar methylation level of every CpG within each sample is observed and the gene follows “on-or-off” methylation pattern. Each circle represents a CpG site in a sample. Area shaded in black is proportional to the methylation level of the CpG site indicated by pyrosequencing. (C) Validation of complete methylation-sensitive restriction enzyme digestion on unmethylated molecules. Genomic DNA was predigested with Bst UI followed by PCR amplification with TUSC3 and ID2 specific primers ( Table S2 ). Bst UI digestion sites within the ID2 region were unmethylated ( Figure S6 ) and, therefore, no PCR product was generated after enzyme digestion. (D) Allele-specific methylation of TUSC3 on the fragment containing SNP rs12550009 demonstrated by enzyme digestion pyrosequencing. The “Simplex” diagrams (top) show the reference pyrograms by genotype. A heterozygous CT in the methylated samples (PM55 and PM123) displays a homozygous T pattern after Bst UI digestion indicating predominant methylation of the T allele. Allele-specific mRNA expression is concordant with allele-specific methylation on the same SNP rs1250009. Predominant expression of C alleles was observed in the cDNAs generated by RNA specific primers (bottom). RT+ and RT− represent assays with Reverse Transcriptase and without Reverse Transcriptase, respectively.
    Bst Ui, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bst ui/product/New England Biolabs
    Average 99 stars, based on 58 article reviews
    Price from $9.99 to $1999.99
    bst ui - by Bioz Stars, 2020-08
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    88
    New England Biolabs bst ui restriction enzyme
    Agarose gel picture illustrating the different <t>A118G</t> genotypes identified by RFLP using the <t>Bst</t> UI restriction enzyme. Column 1 is a 100 bp marker, column 2 shows a sample homozygote for the mutant G118 allele, column 3 shows a sample heterozygote for the A118G SNP and columns 4–8 are samples heterozygous for the wild-type A118 allele
    Bst Ui Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bst ui restriction enzyme/product/New England Biolabs
    Average 88 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    bst ui restriction enzyme - by Bioz Stars, 2020-08
    88/100 stars
      Buy from Supplier

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    Promoter CpG methylation correlates with lack of TUSC3 gene expression. (A) Schematic of TUSC3 locus showing the regions investigated by bisulfite pyrosequencing on the promoter (−105 to −57 relative to the transcriptional start site according to NM_006765) and genotyping assays of the 5′ untranslated region. PCR primers for DNA genotyping are indicated by black arrows while RT-PCR primers for mRNA genotyping are indicated by arrows highlighted in white. Enzyme recognition sites for Bst UI are indicated by “ B ”. (B) Methylation status of TUSC3 promoter region studied by bisulfite pyrosequencing. A similar methylation level of every CpG within each sample is observed and the gene follows “on-or-off” methylation pattern. Each circle represents a CpG site in a sample. Area shaded in black is proportional to the methylation level of the CpG site indicated by pyrosequencing. (C) Validation of complete methylation-sensitive restriction enzyme digestion on unmethylated molecules. Genomic DNA was predigested with Bst UI followed by PCR amplification with TUSC3 and ID2 specific primers ( Table S2 ). Bst UI digestion sites within the ID2 region were unmethylated ( Figure S6 ) and, therefore, no PCR product was generated after enzyme digestion. (D) Allele-specific methylation of TUSC3 on the fragment containing SNP rs12550009 demonstrated by enzyme digestion pyrosequencing. The “Simplex” diagrams (top) show the reference pyrograms by genotype. A heterozygous CT in the methylated samples (PM55 and PM123) displays a homozygous T pattern after Bst UI digestion indicating predominant methylation of the T allele. Allele-specific mRNA expression is concordant with allele-specific methylation on the same SNP rs1250009. Predominant expression of C alleles was observed in the cDNAs generated by RNA specific primers (bottom). RT+ and RT− represent assays with Reverse Transcriptase and without Reverse Transcriptase, respectively.

    Journal: PLoS ONE

    Article Title: Human Placental-Specific Epipolymorphism and its Association with Adverse Pregnancy Outcomes

    doi: 10.1371/journal.pone.0007389

    Figure Lengend Snippet: Promoter CpG methylation correlates with lack of TUSC3 gene expression. (A) Schematic of TUSC3 locus showing the regions investigated by bisulfite pyrosequencing on the promoter (−105 to −57 relative to the transcriptional start site according to NM_006765) and genotyping assays of the 5′ untranslated region. PCR primers for DNA genotyping are indicated by black arrows while RT-PCR primers for mRNA genotyping are indicated by arrows highlighted in white. Enzyme recognition sites for Bst UI are indicated by “ B ”. (B) Methylation status of TUSC3 promoter region studied by bisulfite pyrosequencing. A similar methylation level of every CpG within each sample is observed and the gene follows “on-or-off” methylation pattern. Each circle represents a CpG site in a sample. Area shaded in black is proportional to the methylation level of the CpG site indicated by pyrosequencing. (C) Validation of complete methylation-sensitive restriction enzyme digestion on unmethylated molecules. Genomic DNA was predigested with Bst UI followed by PCR amplification with TUSC3 and ID2 specific primers ( Table S2 ). Bst UI digestion sites within the ID2 region were unmethylated ( Figure S6 ) and, therefore, no PCR product was generated after enzyme digestion. (D) Allele-specific methylation of TUSC3 on the fragment containing SNP rs12550009 demonstrated by enzyme digestion pyrosequencing. The “Simplex” diagrams (top) show the reference pyrograms by genotype. A heterozygous CT in the methylated samples (PM55 and PM123) displays a homozygous T pattern after Bst UI digestion indicating predominant methylation of the T allele. Allele-specific mRNA expression is concordant with allele-specific methylation on the same SNP rs1250009. Predominant expression of C alleles was observed in the cDNAs generated by RNA specific primers (bottom). RT+ and RT− represent assays with Reverse Transcriptase and without Reverse Transcriptase, respectively.

    Article Snippet: For Bst UI predigestion assay followed by pyrosequencing on TUSC3 , 200 ng of genomic DNA was digested with 100 units of Bst UI (New England Biolabs) for 18 hours.

    Techniques: CpG Methylation Assay, Expressing, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Methylation, Amplification, Generated

    Competitive in vitro DNA replication reactions. (A) Competition with 1-246T. For lanes 1 to 3, 1 μg of immunoaffinity-purified T antigen was used to program an in vitro DNA replication reaction of 236 ng of pSV0II+ and 1 μCi of [α- 32 P]dATP in the absence (lanes 1 and 2) or presence (lane 3) of 100 ng of human topo I; lane 1 is a negative control without T antigen. For lanes 4 to 7, T-antigen-mediated in vitro DNA replication was initiated without label for 30 min. Then 1 μCi of [α- 32 P]dATP, 100 ng of human topo I (lanes 5 and 6), and a 10 M excess (over WT T antigen) of 1-246T (lanes 6 and 7) were added, and the reaction continued for an additional 30 min. The labeled DNA was purified, analyzed on a 1.5% agarose gel, dried, and exposed to film. The radioactive products were excised and subjected to Cerenkov counting. (B) Stimulation of completed molecules and overall DNA replication in the presence of topo I (−1-246T) or in the presence of topo I and 1-246T (+1-246T) DNA replication in the presence of T antigen alone is given a value of 1, and fold stimulation is calculated for each molar excess of 1-246T over WT T antigen and depicted graphically. The experiment containing 0.6 M equivalents of 1-246T (relative to WT T antigen) was done with a delayed pulse-chase assay. Experiments containing 2 and 10 M excesses of 1-246T were performed with delayed pulse assays; 10 M excess represents the experiment illustrated in panel A. (C) Restriction enzyme digestion of the form I molecules obtained during a delayed pulse competition reaction. A competition reaction with a 10 M excess of 1-246T was carried out as for panel A. After separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. The 739-bp fragment contains the origin of replication (ori), and the 493- and 330-bp fragments are on the opposite side close to the site of termination. The numbers below the lanes represent relative incorporation into form I DNA. (D) Competition with the catalytic site topo I mutant Y723F. An in vitro DNA replication assay was performed in the presence or absence of various quantities of WT human topo I and mutant topo I Y723F. The DNA was synthesized in the presence of label for 1 h and analyzed on a 1.5% agarose gel. Replicated completed molecules (from CCC to form I on the gel) were quantitated, and the results (from two independent experiments) are expressed as a percentage of the values obtained with WT topo I alone. Samples: 1, no T antigen (negative control); 2, T antigen alone; 3, 100 ng of human topo I; 4, 50 ng of topo I and 50 ng of Y723F; 5, 150 ng of topo I; 6, 50 ng of topo I and 100 ng of Y723F. Samples 1, 2, and 4 were normalized to sample 3, and sample 6 was normalized to sample 5.

    Journal: Molecular and Cellular Biology

    Article Title: Human Topoisomerase I Promotes Initiation of Simian Virus 40 DNA Replication In Vitro

    doi:

    Figure Lengend Snippet: Competitive in vitro DNA replication reactions. (A) Competition with 1-246T. For lanes 1 to 3, 1 μg of immunoaffinity-purified T antigen was used to program an in vitro DNA replication reaction of 236 ng of pSV0II+ and 1 μCi of [α- 32 P]dATP in the absence (lanes 1 and 2) or presence (lane 3) of 100 ng of human topo I; lane 1 is a negative control without T antigen. For lanes 4 to 7, T-antigen-mediated in vitro DNA replication was initiated without label for 30 min. Then 1 μCi of [α- 32 P]dATP, 100 ng of human topo I (lanes 5 and 6), and a 10 M excess (over WT T antigen) of 1-246T (lanes 6 and 7) were added, and the reaction continued for an additional 30 min. The labeled DNA was purified, analyzed on a 1.5% agarose gel, dried, and exposed to film. The radioactive products were excised and subjected to Cerenkov counting. (B) Stimulation of completed molecules and overall DNA replication in the presence of topo I (−1-246T) or in the presence of topo I and 1-246T (+1-246T) DNA replication in the presence of T antigen alone is given a value of 1, and fold stimulation is calculated for each molar excess of 1-246T over WT T antigen and depicted graphically. The experiment containing 0.6 M equivalents of 1-246T (relative to WT T antigen) was done with a delayed pulse-chase assay. Experiments containing 2 and 10 M excesses of 1-246T were performed with delayed pulse assays; 10 M excess represents the experiment illustrated in panel A. (C) Restriction enzyme digestion of the form I molecules obtained during a delayed pulse competition reaction. A competition reaction with a 10 M excess of 1-246T was carried out as for panel A. After separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel. The 739-bp fragment contains the origin of replication (ori), and the 493- and 330-bp fragments are on the opposite side close to the site of termination. The numbers below the lanes represent relative incorporation into form I DNA. (D) Competition with the catalytic site topo I mutant Y723F. An in vitro DNA replication assay was performed in the presence or absence of various quantities of WT human topo I and mutant topo I Y723F. The DNA was synthesized in the presence of label for 1 h and analyzed on a 1.5% agarose gel. Replicated completed molecules (from CCC to form I on the gel) were quantitated, and the results (from two independent experiments) are expressed as a percentage of the values obtained with WT topo I alone. Samples: 1, no T antigen (negative control); 2, T antigen alone; 3, 100 ng of human topo I; 4, 50 ng of topo I and 50 ng of Y723F; 5, 150 ng of topo I; 6, 50 ng of topo I and 100 ng of Y723F. Samples 1, 2, and 4 were normalized to sample 3, and sample 6 was normalized to sample 5.

    Article Snippet: In some cases, after separation of the replication products on a 1.5% agarose gel, the completed molecules were visualized by ethidium bromide, excised from the gel, Cerenkov counted, purified by GeneClean (Bio 101), digested with Bst UI and Ssp I (New England Biolabs), and applied to a 2% agarose gel.

    Techniques: In Vitro, Purification, Negative Control, Labeling, Agarose Gel Electrophoresis, Pulse Chase, Mutagenesis, Synthesized, Countercurrent Chromatography

    The RP2 onshore tandem GAAA repeat is polymorphic in marmosets. Electropherograms of alleles observed in marmosets genotyped via quantitative fluorescent PCR. ( A ) Representative allele profiles from males, which exhibited only the major allele, and female animals with three distinct genotypes (homozygotes for either the minor or major allele or heterozygotes) are shown. ( B ) Representative random XCI pattern observed at the 5 me CpG-sensitive Bst UI recognition site within the RP2 GAAA-containing amplimer, with an Xa/Xi ratio of approximately 65%. The allele names are the lengths in base pairs of each fluorescence peak and the intensity of each peak is in relative fluorescence units (RFU).

    Journal: PLoS ONE

    Article Title: 5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation

    doi: 10.1371/journal.pone.0103714

    Figure Lengend Snippet: The RP2 onshore tandem GAAA repeat is polymorphic in marmosets. Electropherograms of alleles observed in marmosets genotyped via quantitative fluorescent PCR. ( A ) Representative allele profiles from males, which exhibited only the major allele, and female animals with three distinct genotypes (homozygotes for either the minor or major allele or heterozygotes) are shown. ( B ) Representative random XCI pattern observed at the 5 me CpG-sensitive Bst UI recognition site within the RP2 GAAA-containing amplimer, with an Xa/Xi ratio of approximately 65%. The allele names are the lengths in base pairs of each fluorescence peak and the intensity of each peak is in relative fluorescence units (RFU).

    Article Snippet: This result may be related to the fact that in this particular target sequence the Hha I site overlaps the CpG within the Bst UI site and that overlapping CpG sites may block or impair cleavage if methylated (New England Biolabs usage guidelines).

    Techniques: Polymerase Chain Reaction, Fluorescence

    Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).

    Journal: PLoS ONE

    Article Title: 5meCpG Epigenetic Marks Neighboring a Primate-Conserved Core Promoter Short Tandem Repeat Indicate X-Chromosome Inactivation

    doi: 10.1371/journal.pone.0103714

    Figure Lengend Snippet: Methylation statuses at CpG sites near the RP2 onshore tandem GAAA repeat. Random ( A ) and non-random ( B ) X-inactivation patterns generated for different CpG-containing 5 me CpG-sensitive restriction endonuclease sites obtained using the 5 me CpG-based PCR RP2 / AR biplex assay across the restriction sites. Electropherograms of alleles observed in either undigested genomic DNA or DNA digested with Hpa II, Hha I or Bst UI from females genotyped via quantitative fluorescent PCR are shown. The boxed numbers correspond to the areas under the allele peaks and the intensity of each peak is in relative fluorescence units (RFU).

    Article Snippet: This result may be related to the fact that in this particular target sequence the Hha I site overlaps the CpG within the Bst UI site and that overlapping CpG sites may block or impair cleavage if methylated (New England Biolabs usage guidelines).

    Techniques: Methylation, Generated, Polymerase Chain Reaction, Fluorescence

    Agarose gel picture illustrating the different A118G genotypes identified by RFLP using the Bst UI restriction enzyme. Column 1 is a 100 bp marker, column 2 shows a sample homozygote for the mutant G118 allele, column 3 shows a sample heterozygote for the A118G SNP and columns 4–8 are samples heterozygous for the wild-type A118 allele

    Journal: The Journal of Headache and Pain

    Article Title: The human ?-opioid receptor gene polymorphism (A118G) is associated with head pain severity in a clinical cohort of female migraine with aura patients

    doi: 10.1007/s10194-012-0468-z

    Figure Lengend Snippet: Agarose gel picture illustrating the different A118G genotypes identified by RFLP using the Bst UI restriction enzyme. Column 1 is a 100 bp marker, column 2 shows a sample homozygote for the mutant G118 allele, column 3 shows a sample heterozygote for the A118G SNP and columns 4–8 are samples heterozygous for the wild-type A118 allele

    Article Snippet: Genotyping for the A118G polymorphism was performed by digesting PCR products with the Bst UI restriction enzyme (New England Biolabs, Beverly, MA, USA).

    Techniques: Agarose Gel Electrophoresis, Marker, Mutagenesis