polymerase chain reaction restriction fragment length polymorphism  (Bio-Rad)

 
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    Bio-Rad polymerase chain reaction restriction fragment length polymorphism
    Polymerase Chain Reaction Restriction Fragment Length Polymorphism, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Amplification:

    Article Title: Relationship between Job Stress and 5-HT2A Receptor Polymorphisms on Self-Reported Sleep Quality in Physicians in Urumqi (Xinjiang, China): A Cross-Sectional Study
    Article Snippet: .. The T102C and -1438G/A polymorphisms were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). gDNA (100 ng) was used in each reaction mixture, with the final volume of each reaction mixture totaling 20 μL. gDNA was then amplified using the PCR instrument (MyCycler, Bio-Rad, Hercules, CA, USA). ..

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    Agarose Gel Electrophoresis:

    Article Title: Decreased prevalence of Plasmodium falciparum resistance markers to amodiaquine despite its wide scale use as ACT partner drug in Zanzibar
    Article Snippet: .. PCR-RFLP products were visualized under UV transillumination (GelDoc 2000, BioRad, Hercules® , CA, USA) after 2–2.5% agarose gel electrophoresis and ethidium-bromide staining. .. A mixed infection was considered to contain two P. falciparum strains, contributing with one of each SNP alleles during PCR-RFLP.

    Fluorescence:

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    Ligation:

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    Electrophoresis:

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    Polymerase Chain Reaction:

    Article Title: Polymorphism of FABP2 and PPARG2 genes in risk prediction of cataract among North Indian population
    Article Snippet: .. PPARG2 polymorphism Genotyping was performed using PCR-RFLP (MJ Mini Thermo Cycler-BioRad). with the following primers: forward primer 5′-CAA GCC CAG TCC TTT CTG TG-3′ and reverse primer 5′-AGT GAA GGA ATC GCT TTC CG-3′ ( ). .. All reactions were performed in a total volume of 50 μl containing 10 mmol/l Tris–HCl (pH 8.8), 50 mmol/l KCl, 1.5 mmol/l MgCl2 , 0.2 mmol of each dNTPs, 50 pmol of each of the two primers, 0.3 U of Taq DNA polymerase (Bioline Ltd., London, UK) and 200 ng of genomic DNA.

    Article Title: Relationship between Job Stress and 5-HT2A Receptor Polymorphisms on Self-Reported Sleep Quality in Physicians in Urumqi (Xinjiang, China): A Cross-Sectional Study
    Article Snippet: .. The T102C and -1438G/A polymorphisms were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). gDNA (100 ng) was used in each reaction mixture, with the final volume of each reaction mixture totaling 20 μL. gDNA was then amplified using the PCR instrument (MyCycler, Bio-Rad, Hercules, CA, USA). ..

    Article Title: CTLA4 +49AG (rs231775) and CT60 (rs3087243) gene variants are not associated with alopecia areata in a Mexican population from Monterrey Mexico
    Article Snippet: .. Genotype and allele frequencies of the CTLA4 +49AG (rs231775) andCT60 (rs3087243) gene variants were characterized by PCR-RFLP using a MJ Mini PTC1148 thermal cycler (Bio-Rad, Hercules; CA, USA), specific oligonucleotide primers (IDT, Coralville, IA, USA) and restriction enzymes (New England Biolabs, Ipswich, MA, USA) according to a previously published protocol. .. The fragments obtained were analyzed by electrophoresis in a 2.5% agarose gel containing ethidium bromide, visualized in UVP model 2UV High Performance Transilluminator (Upland, CA, USA) and documented.

    Article Title: Decreased prevalence of Plasmodium falciparum resistance markers to amodiaquine despite its wide scale use as ACT partner drug in Zanzibar
    Article Snippet: .. PCR-RFLP products were visualized under UV transillumination (GelDoc 2000, BioRad, Hercules® , CA, USA) after 2–2.5% agarose gel electrophoresis and ethidium-bromide staining. .. A mixed infection was considered to contain two P. falciparum strains, contributing with one of each SNP alleles during PCR-RFLP.

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    CTG Assay:

    Article Title: Polymorphism of FABP2 and PPARG2 genes in risk prediction of cataract among North Indian population
    Article Snippet: .. PPARG2 polymorphism Genotyping was performed using PCR-RFLP (MJ Mini Thermo Cycler-BioRad). with the following primers: forward primer 5′-CAA GCC CAG TCC TTT CTG TG-3′ and reverse primer 5′-AGT GAA GGA ATC GCT TTC CG-3′ ( ). .. All reactions were performed in a total volume of 50 μl containing 10 mmol/l Tris–HCl (pH 8.8), 50 mmol/l KCl, 1.5 mmol/l MgCl2 , 0.2 mmol of each dNTPs, 50 pmol of each of the two primers, 0.3 U of Taq DNA polymerase (Bioline Ltd., London, UK) and 200 ng of genomic DNA.

    Staining:

    Article Title: Decreased prevalence of Plasmodium falciparum resistance markers to amodiaquine despite its wide scale use as ACT partner drug in Zanzibar
    Article Snippet: .. PCR-RFLP products were visualized under UV transillumination (GelDoc 2000, BioRad, Hercules® , CA, USA) after 2–2.5% agarose gel electrophoresis and ethidium-bromide staining. .. A mixed infection was considered to contain two P. falciparum strains, contributing with one of each SNP alleles during PCR-RFLP.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

    Allele-specific Oligonucleotide:

    Article Title: Polymorphisms in genes involved in drug detoxification and clinical outcomes of anthracycline-based neoadjuvant chemotherapy in Chinese Han breast cancer patients
    Article Snippet: .. The assay was implemented via three steps: a triplex PCR that resulted in the amplified target sequences with MnSOD T47C, CAT C-262T and GSTP1 A313G loci, a triplex allele-specific oligonucleotide probe ligation reaction that generates the ligation between perfectly matched probes at their junctions while no ligation occurred between mismatched ones and an analysis of genotype-specific ligation products by both fluorescence and size after electrophoresis on ABI PRISM 377 DNA sequencer., MDR1 C3435T, G2677T/A and C1236T polymorphisms were analyzed by PCR-RFLP as described by Ni et al. PCR reaction was performed on a DNA thermal cycler (Bio-Rad iCycle). .. Controls (no DNA template) were run to ensure there was no amplification of contaminating DNA, and Beta-actin was run as internal control.

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    Bio-Rad rflp
    Hae <t>III</t> enzyme digestion identification of β-fibrinogen -148C/T (polymerase chain reaction and restriction fragment length polymorphism). M: Marker; C/C: 202 bp, 98 bp; C/T: 300 bp, 202 bp, 98 bp; T/T: 300 bp.
    Rflp, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad pcr rflp
    Agarose gel picture showing <t>PCR-RFLP</t> product of PPARγ2 gene, lane 1 shows undigested product, lanes 2, 3, 4, 5, 7, 8, 9, 10, 11, and 12 show CG(+/-) genotype and lane 6 shows 100 bp ladder.
    Pcr Rflp, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad pcr restriction fragment length polymorphism rflp analysis gels
    Screening for A2058G and A2059G mutations in B. pertussis by <t>PCR-RFLP</t> analysis. (A). Bsa I (lanes 1 to 5) or Bbs I (lanes 6 to 10) digestion of a 521-bp fragment of the 23S rDNA gene of erythromycin-resistant B. pertussis clinical isolates (A228, C353, and MN2531), heterogeneous strain C352, and erythromycin-susceptible strain MN2726. The 521-bp fragment was generated by PCR amplification using primers 1907U and 2408L as described in Materials and Methods. Lanes: M, 100-bp ladder (Life Technologies); 1 and 6, B. pertussis A228; 2 and 7, B. pertussis C352; 3 and 8, B. pertussis C353; 4 and 9, B. pertussis MN2531; 5 and 10, B. pertussis MN2726. (B). Bbs I digestion of the 521-bp fragment of additional isolates of B. pertussis . Lanes: M, 100-bp ladder (Life Technologies); 1 to 7, erythromycin-susceptible clinical isolates B. pertussis MN277, MN973, MN1286, MN1699, MN1773, MN1893, and MN2726; 8, erythromycin-resistant B. pertussis isolate MN253.
    Pcr Restriction Fragment Length Polymorphism Rflp Analysis Gels, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Hae III enzyme digestion identification of β-fibrinogen -148C/T (polymerase chain reaction and restriction fragment length polymorphism). M: Marker; C/C: 202 bp, 98 bp; C/T: 300 bp, 202 bp, 98 bp; T/T: 300 bp.

    Journal: Neural Regeneration Research

    Article Title: Relationship between the -455G/A and -148C/T polymorphisms in the beta-fibrinogen gene and cerebral infarction in the Xinjiang Uygur and Han Chinese populations ☆

    doi: 10.3969/j.issn.1673-5374.2012.07.012

    Figure Lengend Snippet: Hae III enzyme digestion identification of β-fibrinogen -148C/T (polymerase chain reaction and restriction fragment length polymorphism). M: Marker; C/C: 202 bp, 98 bp; C/T: 300 bp, 202 bp, 98 bp; T/T: 300 bp.

    Article Snippet: Both amplified products included a Hae III (New England Biolabs, Ipswich, MA, USA) restriction site, forming an RFLP (Bio-Rad, Hercules, CA, USA).

    Techniques: Polymerase Chain Reaction, Marker

    Hae III enzyme digestion identification of β-fibrinogen -455G/A (polymerase chain reaction and restriction fragment length polymorphism). M: DL2000 marker; 1: AA (669 bp); 2, 6, 7, 8: GG (488 bp, 181 bp); 3–5: GA (455 bp).

    Journal: Neural Regeneration Research

    Article Title: Relationship between the -455G/A and -148C/T polymorphisms in the beta-fibrinogen gene and cerebral infarction in the Xinjiang Uygur and Han Chinese populations ☆

    doi: 10.3969/j.issn.1673-5374.2012.07.012

    Figure Lengend Snippet: Hae III enzyme digestion identification of β-fibrinogen -455G/A (polymerase chain reaction and restriction fragment length polymorphism). M: DL2000 marker; 1: AA (669 bp); 2, 6, 7, 8: GG (488 bp, 181 bp); 3–5: GA (455 bp).

    Article Snippet: Both amplified products included a Hae III (New England Biolabs, Ipswich, MA, USA) restriction site, forming an RFLP (Bio-Rad, Hercules, CA, USA).

    Techniques: Polymerase Chain Reaction, Marker

    Fluorescent protein-2A-ZFN constructs enable efficient knockin genome editing in demanding applications. ( A ) K562 or Jurkat cells were transfected with mRNA expressing EGFP-2A-ZFNL and DsRed2–2A-ZFNR for RSK4 along with an ssODN knockin donor ( RSK4 Cys443Val/BamHI). For K562, the cells were diluted 3 days post-transfection with mock-transfected cells to produce a pool of ∼0.5% ZFN-expressing cells. Cells of this pool were seeded singly or subjected to FACS isolation of EGFP/DsRed2 double-fluorescent cells (any level) that were thereafter also seeded singly. For Jurkat, non-sorted cells or cells FACS isolated from the 15% most highly EGFP/DsRed2 double-fluorescent cell population were seeded singly 3 days post-transfection. In both experiments, the singly seeded cells were expanded to clonal cell lines and analysed for knockin mutation by RFLP analysis. For K562, data are means + range of two independent experiments. ( B ) K562 cells were co-transfected with mRNA expressing EGFP-2A-ZFNs (left and right) for RSK4 and DsRed2–2A-ZFNs (left and right) for PRMT1 along with ssODN donors for knockin modification of these loci ( RSK4 Cys443Val/BamHI and PRMT1 ScaI). Three days after transfection, non-sorted cells or cells FACS isolated from the 15% most highly EGFP/DsRed2 double-fluorescent cell population were seeded singly, expanded to clonal cell lines and analysed for complete knockin modification of the RSK4 and PRMT1 loci. Data are summed of three independent experiments. ( C ) Overview of strategy for generation of cell pools with high levels of stable knockin modification. In this case, left and right ZFNs of a ZFN pair (ZFNL and ZFNR) have been coupled to distinct fluorescent proteins. ( D ) Example of close to 100% knockin in an MCF10A cell pool. Specifically, MCF10A cells were transfected for codon-conversion knockin at the RSK2 locus as described in Figure 1E , (ii). Three days after transfection, the 40% most highly fluorescent cells were FACS isolated and cultured as a pool for 2 weeks. Thereafter, this cell pool was subjected to the same treatment for two more rounds. After the third round, the cell pool was analysed for stable knockin at the RSK2 locus by RFLP analysis with untreated MCF10A cells serving as a control. PCR products derived from wild-type and mutant alleles are indicated by an asterisk and arrows, respectively.

    Journal: Nucleic Acids Research

    Article Title: High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs

    doi: 10.1093/nar/gku251

    Figure Lengend Snippet: Fluorescent protein-2A-ZFN constructs enable efficient knockin genome editing in demanding applications. ( A ) K562 or Jurkat cells were transfected with mRNA expressing EGFP-2A-ZFNL and DsRed2–2A-ZFNR for RSK4 along with an ssODN knockin donor ( RSK4 Cys443Val/BamHI). For K562, the cells were diluted 3 days post-transfection with mock-transfected cells to produce a pool of ∼0.5% ZFN-expressing cells. Cells of this pool were seeded singly or subjected to FACS isolation of EGFP/DsRed2 double-fluorescent cells (any level) that were thereafter also seeded singly. For Jurkat, non-sorted cells or cells FACS isolated from the 15% most highly EGFP/DsRed2 double-fluorescent cell population were seeded singly 3 days post-transfection. In both experiments, the singly seeded cells were expanded to clonal cell lines and analysed for knockin mutation by RFLP analysis. For K562, data are means + range of two independent experiments. ( B ) K562 cells were co-transfected with mRNA expressing EGFP-2A-ZFNs (left and right) for RSK4 and DsRed2–2A-ZFNs (left and right) for PRMT1 along with ssODN donors for knockin modification of these loci ( RSK4 Cys443Val/BamHI and PRMT1 ScaI). Three days after transfection, non-sorted cells or cells FACS isolated from the 15% most highly EGFP/DsRed2 double-fluorescent cell population were seeded singly, expanded to clonal cell lines and analysed for complete knockin modification of the RSK4 and PRMT1 loci. Data are summed of three independent experiments. ( C ) Overview of strategy for generation of cell pools with high levels of stable knockin modification. In this case, left and right ZFNs of a ZFN pair (ZFNL and ZFNR) have been coupled to distinct fluorescent proteins. ( D ) Example of close to 100% knockin in an MCF10A cell pool. Specifically, MCF10A cells were transfected for codon-conversion knockin at the RSK2 locus as described in Figure 1E , (ii). Three days after transfection, the 40% most highly fluorescent cells were FACS isolated and cultured as a pool for 2 weeks. Thereafter, this cell pool was subjected to the same treatment for two more rounds. After the third round, the cell pool was analysed for stable knockin at the RSK2 locus by RFLP analysis with untreated MCF10A cells serving as a control. PCR products derived from wild-type and mutant alleles are indicated by an asterisk and arrows, respectively.

    Article Snippet: Densitometric quantization of CEL-I and RFLP assays Gels with digested PCR products were stained for 10 min with ethidium bromide and imaged with a Quantity One Gel Doc XR imaging system (Bio-Rad).

    Techniques: Construct, Knock-In, Transfection, Expressing, FACS, Isolation, Mutagenesis, Modification, Cell Culture, Polymerase Chain Reaction, Derivative Assay

    CRISPR/Cas9 genome editing is enhanced using the 2A-coupled fluorescent protein/FACS strategy. ( A ) Schematic of a construct, pCMV-Cas9-FP, that co-expresses gRNA and Cas6–2A-fluorescent protein. Functional parts indicated are U6 promoter, guide (g)RNA, CMV promoter, Cas9 nuclease, 2A peptide and fluorescent protein (GFP). ( B ) – ( D ) K562 cells were transfected (B) with a Cas9-GFP construct targeting the KRAS locus, (C) with a Cas9-GFP construct targeting the VEGFA locus or the same construct containing a nickase version of Cas9 (Cas9N) or (D) with such constructs targeting the EMX1 locus as a nickase pair or as individual nickases coupled to GFP or dsRed2. An ssODN donor ( EMX1 gRNA 1/2 EcoRV) for knockin of an EcoRV restriction site in EMX1 was co-transfected. Three days after transfection, genomic DNA was isolated from non-sorted cells or from cells FACS isolated for low, medium or high fluorescence intensities and analysed by RFLP or CEL-I assay, as indicated. In all experiments, repeated three times with similar results, PCR products containing amplicons derived from mutant alleles or from wild-type alleles only are indicated by the arrows and asterisks, respectively. N.D.: none detected.

    Journal: Nucleic Acids Research

    Article Title: High-efficiency genome editing via 2A-coupled co-expression of fluorescent proteins and zinc finger nucleases or CRISPR/Cas9 nickase pairs

    doi: 10.1093/nar/gku251

    Figure Lengend Snippet: CRISPR/Cas9 genome editing is enhanced using the 2A-coupled fluorescent protein/FACS strategy. ( A ) Schematic of a construct, pCMV-Cas9-FP, that co-expresses gRNA and Cas6–2A-fluorescent protein. Functional parts indicated are U6 promoter, guide (g)RNA, CMV promoter, Cas9 nuclease, 2A peptide and fluorescent protein (GFP). ( B ) – ( D ) K562 cells were transfected (B) with a Cas9-GFP construct targeting the KRAS locus, (C) with a Cas9-GFP construct targeting the VEGFA locus or the same construct containing a nickase version of Cas9 (Cas9N) or (D) with such constructs targeting the EMX1 locus as a nickase pair or as individual nickases coupled to GFP or dsRed2. An ssODN donor ( EMX1 gRNA 1/2 EcoRV) for knockin of an EcoRV restriction site in EMX1 was co-transfected. Three days after transfection, genomic DNA was isolated from non-sorted cells or from cells FACS isolated for low, medium or high fluorescence intensities and analysed by RFLP or CEL-I assay, as indicated. In all experiments, repeated three times with similar results, PCR products containing amplicons derived from mutant alleles or from wild-type alleles only are indicated by the arrows and asterisks, respectively. N.D.: none detected.

    Article Snippet: Densitometric quantization of CEL-I and RFLP assays Gels with digested PCR products were stained for 10 min with ethidium bromide and imaged with a Quantity One Gel Doc XR imaging system (Bio-Rad).

    Techniques: CRISPR, FACS, Construct, Functional Assay, Transfection, Knock-In, Isolation, Fluorescence, Polymerase Chain Reaction, Derivative Assay, Mutagenesis

    Agarose gel picture showing PCR-RFLP product of PPARγ2 gene, lane 1 shows undigested product, lanes 2, 3, 4, 5, 7, 8, 9, 10, 11, and 12 show CG(+/-) genotype and lane 6 shows 100 bp ladder.

    Journal: Meta Gene

    Article Title: Polymorphism of FABP2 and PPARG2 genes in risk prediction of cataract among North Indian population

    doi: 10.1016/j.mgene.2014.02.002

    Figure Lengend Snippet: Agarose gel picture showing PCR-RFLP product of PPARγ2 gene, lane 1 shows undigested product, lanes 2, 3, 4, 5, 7, 8, 9, 10, 11, and 12 show CG(+/-) genotype and lane 6 shows 100 bp ladder.

    Article Snippet: PPARG2 polymorphism Genotyping was performed using PCR-RFLP (MJ Mini Thermo Cycler-BioRad). with the following primers: forward primer 5′-CAA GCC CAG TCC TTT CTG TG-3′ and reverse primer 5′-AGT GAA GGA ATC GCT TTC CG-3′ ( ).

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction

    Screening for A2058G and A2059G mutations in B. pertussis by PCR-RFLP analysis. (A). Bsa I (lanes 1 to 5) or Bbs I (lanes 6 to 10) digestion of a 521-bp fragment of the 23S rDNA gene of erythromycin-resistant B. pertussis clinical isolates (A228, C353, and MN2531), heterogeneous strain C352, and erythromycin-susceptible strain MN2726. The 521-bp fragment was generated by PCR amplification using primers 1907U and 2408L as described in Materials and Methods. Lanes: M, 100-bp ladder (Life Technologies); 1 and 6, B. pertussis A228; 2 and 7, B. pertussis C352; 3 and 8, B. pertussis C353; 4 and 9, B. pertussis MN2531; 5 and 10, B. pertussis MN2726. (B). Bbs I digestion of the 521-bp fragment of additional isolates of B. pertussis . Lanes: M, 100-bp ladder (Life Technologies); 1 to 7, erythromycin-susceptible clinical isolates B. pertussis MN277, MN973, MN1286, MN1699, MN1773, MN1893, and MN2726; 8, erythromycin-resistant B. pertussis isolate MN253.

    Journal: Journal of Clinical Microbiology

    Article Title: Identification of a Mutation Associated with Erythromycin Resistance in Bordetella pertussis: Implications for Surveillance of Antimicrobial Resistance

    doi: 10.1128/JCM.41.3.1167-1172.2003

    Figure Lengend Snippet: Screening for A2058G and A2059G mutations in B. pertussis by PCR-RFLP analysis. (A). Bsa I (lanes 1 to 5) or Bbs I (lanes 6 to 10) digestion of a 521-bp fragment of the 23S rDNA gene of erythromycin-resistant B. pertussis clinical isolates (A228, C353, and MN2531), heterogeneous strain C352, and erythromycin-susceptible strain MN2726. The 521-bp fragment was generated by PCR amplification using primers 1907U and 2408L as described in Materials and Methods. Lanes: M, 100-bp ladder (Life Technologies); 1 and 6, B. pertussis A228; 2 and 7, B. pertussis C352; 3 and 8, B. pertussis C353; 4 and 9, B. pertussis MN2531; 5 and 10, B. pertussis MN2726. (B). Bbs I digestion of the 521-bp fragment of additional isolates of B. pertussis . Lanes: M, 100-bp ladder (Life Technologies); 1 to 7, erythromycin-susceptible clinical isolates B. pertussis MN277, MN973, MN1286, MN1699, MN1773, MN1893, and MN2726; 8, erythromycin-resistant B. pertussis isolate MN253.

    Article Snippet: Quantification of amplicon restriction fragments on PCR-restriction fragment length polymorphism (RFLP) analysis gels was done by densitometry using Quantity One software (Bio-Rad).

    Techniques: Polymerase Chain Reaction, Generated, Amplification