polymerase chain reaction pcr genomic dna  (Roche)

 
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    Roche polymerase chain reaction pcr genomic dna
    Polymerase Chain Reaction Pcr Genomic Dna, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction pcr genomic dna/product/Roche
    Average 92 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction pcr genomic dna - by Bioz Stars, 2020-05
    92/100 stars

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    DNA Extraction:

    Article Title: High frequency of mutations in gyrA gene associated with quinolones resistance in uropathogenic Escherichiacoli isolates from the north of Iran
    Article Snippet: .. DNA extraction and polymerase chain reaction (PCR) Genomic DNA of all isolates was extracted using High pure DNA template preparation kit (Roche, Germany) as stated by manufacturer instruction. .. PCR reaction was performed to amplify gyrA gene in Quinolone resistant determining region (QRDR) using specific primers, gyrA -F: 5’-GCT GCC AGA TGT CCG AGA T-3’, gyrA -R: 5’-TCC GTG CCG TCA TAG TTA TCA-3’.

    Polymerase Chain Reaction:

    Article Title: High frequency of mutations in gyrA gene associated with quinolones resistance in uropathogenic Escherichiacoli isolates from the north of Iran
    Article Snippet: .. DNA extraction and polymerase chain reaction (PCR) Genomic DNA of all isolates was extracted using High pure DNA template preparation kit (Roche, Germany) as stated by manufacturer instruction. .. PCR reaction was performed to amplify gyrA gene in Quinolone resistant determining region (QRDR) using specific primers, gyrA -F: 5’-GCT GCC AGA TGT CCG AGA T-3’, gyrA -R: 5’-TCC GTG CCG TCA TAG TTA TCA-3’.

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    Roche bisulfite pyrosequencing genomic dna
    Lentiviral-mediated Kaiso knockdown (Kaiso KD ) is associated with a reduction of ICR1 methylation and decreased endogenous IGF2 and H19 transcription in human primary. As controls, transfections with shRNA against a scrambled locus ( S.L. ) were performed. a Knockdown efficiency was quantitated by RT-qPCR. The expression values are normalized to PDH and B2M transcripts and are represented as mean ± SEM from two independent experiments each. p values for the transcriptional comparisons are indicated. b ICR1 methylation was determined by <t>pyrosequencing</t> of the three established differentially methylated ICR1 CpGs in bisulfite-treated <t>DNA.</t> The location of the analysed CpGs is schematically depicted on top . Mean methylation values of three replicates are presented ± SEM for each CG position. The knockdown of Kaiso is associated with reduced methylation of endogenous ICR1
    Bisulfite Pyrosequencing Genomic Dna, supplied by Roche, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bisulfite pyrosequencing genomic dna/product/Roche
    Average 90 stars, based on 1 article reviews
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    92
    Roche hekn genomic dna
    ( A ) Regulation of Jun B mRNA by DHA. <t>HEKn</t> cells were treated with ethanol vehicle (control), or with either 10 μM DHA or 20 μM DHA. RNA isolation, synthesis of first strand <t>DNA,</t> and real-time PCR are described in Methods. Results are means from three independent experiments ± STDEV. The double asterisk (**) denotes an average that is statistically significant by Student’s t -test from the ethanol control, p
    Hekn Genomic Dna, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hekn genomic dna/product/Roche
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    92
    Roche genomic dna
    GRN played a pivotal role in the <t>ALD-DNA-induced</t> M2b polarization in vitro . (A–B) Primary macrophages were stimulated with increasing amounts of ALD-DNA for 24 h. mRNA levels of GRN in macrophages were analyzed by real time <t>PCR</t> analysis (A), and protein levels of GRN in the supernatants of macrophages were analyzed by western blot (B). Above, quantitative results of western blots, the band intensity was measured by Image J; Below, representative western blots. Similar results were obtained in three independent experiments. Data are representative of results obtained in three independent experiments. Primary peritoneal macrophages were stimulated by ALD-DNA (50 µg/mL) with GRN (5 µg/mL) for 24 h (C and F), or were pretreated with elastase inhibitor (100 µM) or DMSO (0.1%) for 12 h (D and G), and then were exposed to ALD-DNA, UnALD-DNA, or PBS for another 24 h. Macrophages were transfected with control siRNA (200 nM) or GRN siRNA (siGRN, 200 nM). 36 h posttransfection, macrophages were stimulated with PBS, UnALD-DNA or ALD-DNA (50 µg/mL) (E and H). (C–E) ELISA assay was used to analyze the levels of TNF-α, IL-1β, IL-6, IL-10, IL-12, and MCP-1 in the culture supernatants of macrophages. Data are means ± SD of three independent experiments. (F–H) Western blot analysis was used to analyze the protein levels of iNOS in macrophages. Data are representative of three separate experiments. Similar results were obtained in three independent experiments. Band intensity was measured by Image J and the ratios of iNOS to β-actin were calculated. *, P
    Genomic Dna, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Lentiviral-mediated Kaiso knockdown (Kaiso KD ) is associated with a reduction of ICR1 methylation and decreased endogenous IGF2 and H19 transcription in human primary. As controls, transfections with shRNA against a scrambled locus ( S.L. ) were performed. a Knockdown efficiency was quantitated by RT-qPCR. The expression values are normalized to PDH and B2M transcripts and are represented as mean ± SEM from two independent experiments each. p values for the transcriptional comparisons are indicated. b ICR1 methylation was determined by pyrosequencing of the three established differentially methylated ICR1 CpGs in bisulfite-treated DNA. The location of the analysed CpGs is schematically depicted on top . Mean methylation values of three replicates are presented ± SEM for each CG position. The knockdown of Kaiso is associated with reduced methylation of endogenous ICR1

    Journal: Clinical Epigenetics

    Article Title: Kaiso mediates human ICR1 methylation maintenance and H19 transcriptional fine regulation

    doi: 10.1186/s13148-016-0215-4

    Figure Lengend Snippet: Lentiviral-mediated Kaiso knockdown (Kaiso KD ) is associated with a reduction of ICR1 methylation and decreased endogenous IGF2 and H19 transcription in human primary. As controls, transfections with shRNA against a scrambled locus ( S.L. ) were performed. a Knockdown efficiency was quantitated by RT-qPCR. The expression values are normalized to PDH and B2M transcripts and are represented as mean ± SEM from two independent experiments each. p values for the transcriptional comparisons are indicated. b ICR1 methylation was determined by pyrosequencing of the three established differentially methylated ICR1 CpGs in bisulfite-treated DNA. The location of the analysed CpGs is schematically depicted on top . Mean methylation values of three replicates are presented ± SEM for each CG position. The knockdown of Kaiso is associated with reduced methylation of endogenous ICR1

    Article Snippet: Bisulfite pyrosequencing Genomic DNA of primary fibroblasts was extracted with the High Pure PCR Template preparation Kit (Roche) according to the manufacturer’s instructions.

    Techniques: Methylation, Transfection, shRNA, Quantitative RT-PCR, Expressing

    ( A ) Regulation of Jun B mRNA by DHA. HEKn cells were treated with ethanol vehicle (control), or with either 10 μM DHA or 20 μM DHA. RNA isolation, synthesis of first strand DNA, and real-time PCR are described in Methods. Results are means from three independent experiments ± STDEV. The double asterisk (**) denotes an average that is statistically significant by Student’s t -test from the ethanol control, p

    Journal: Nutrients

    Article Title: Bioactive Dietary VDR Ligands Regulate Genes Encoding Biomarkers of Skin Repair That Are Associated with Risk for Psoriasis

    doi: 10.3390/nu10020174

    Figure Lengend Snippet: ( A ) Regulation of Jun B mRNA by DHA. HEKn cells were treated with ethanol vehicle (control), or with either 10 μM DHA or 20 μM DHA. RNA isolation, synthesis of first strand DNA, and real-time PCR are described in Methods. Results are means from three independent experiments ± STDEV. The double asterisk (**) denotes an average that is statistically significant by Student’s t -test from the ethanol control, p

    Article Snippet: PCR reactions contained approximately 130 ng/μL of HEKn genomic DNA from a single lot of cells, 0.5 μL of an 18 μM stock of the above three primers (0.9 μM final concentration of each primer), and 5 μL of Fast Start Universal SYBR Green Master Mix from Roche Applied Science (Indianapolis, IN, USA) in a total volume of 10 μL.

    Techniques: Isolation, Real-time Polymerase Chain Reaction

    GRN played a pivotal role in the ALD-DNA-induced M2b polarization in vitro . (A–B) Primary macrophages were stimulated with increasing amounts of ALD-DNA for 24 h. mRNA levels of GRN in macrophages were analyzed by real time PCR analysis (A), and protein levels of GRN in the supernatants of macrophages were analyzed by western blot (B). Above, quantitative results of western blots, the band intensity was measured by Image J; Below, representative western blots. Similar results were obtained in three independent experiments. Data are representative of results obtained in three independent experiments. Primary peritoneal macrophages were stimulated by ALD-DNA (50 µg/mL) with GRN (5 µg/mL) for 24 h (C and F), or were pretreated with elastase inhibitor (100 µM) or DMSO (0.1%) for 12 h (D and G), and then were exposed to ALD-DNA, UnALD-DNA, or PBS for another 24 h. Macrophages were transfected with control siRNA (200 nM) or GRN siRNA (siGRN, 200 nM). 36 h posttransfection, macrophages were stimulated with PBS, UnALD-DNA or ALD-DNA (50 µg/mL) (E and H). (C–E) ELISA assay was used to analyze the levels of TNF-α, IL-1β, IL-6, IL-10, IL-12, and MCP-1 in the culture supernatants of macrophages. Data are means ± SD of three independent experiments. (F–H) Western blot analysis was used to analyze the protein levels of iNOS in macrophages. Data are representative of three separate experiments. Similar results were obtained in three independent experiments. Band intensity was measured by Image J and the ratios of iNOS to β-actin were calculated. *, P

    Journal: PLoS ONE

    Article Title: Granulin Exacerbates Lupus Nephritis via Enhancing Macrophage M2b Polarization

    doi: 10.1371/journal.pone.0065542

    Figure Lengend Snippet: GRN played a pivotal role in the ALD-DNA-induced M2b polarization in vitro . (A–B) Primary macrophages were stimulated with increasing amounts of ALD-DNA for 24 h. mRNA levels of GRN in macrophages were analyzed by real time PCR analysis (A), and protein levels of GRN in the supernatants of macrophages were analyzed by western blot (B). Above, quantitative results of western blots, the band intensity was measured by Image J; Below, representative western blots. Similar results were obtained in three independent experiments. Data are representative of results obtained in three independent experiments. Primary peritoneal macrophages were stimulated by ALD-DNA (50 µg/mL) with GRN (5 µg/mL) for 24 h (C and F), or were pretreated with elastase inhibitor (100 µM) or DMSO (0.1%) for 12 h (D and G), and then were exposed to ALD-DNA, UnALD-DNA, or PBS for another 24 h. Macrophages were transfected with control siRNA (200 nM) or GRN siRNA (siGRN, 200 nM). 36 h posttransfection, macrophages were stimulated with PBS, UnALD-DNA or ALD-DNA (50 µg/mL) (E and H). (C–E) ELISA assay was used to analyze the levels of TNF-α, IL-1β, IL-6, IL-10, IL-12, and MCP-1 in the culture supernatants of macrophages. Data are means ± SD of three independent experiments. (F–H) Western blot analysis was used to analyze the protein levels of iNOS in macrophages. Data are representative of three separate experiments. Similar results were obtained in three independent experiments. Band intensity was measured by Image J and the ratios of iNOS to β-actin were calculated. *, P

    Article Snippet: Genomic DNA was subjected to quantitative real-time PCR using a Lightcycler480 and SYBR Green system (Roche Diagnostic Systems) following the manufacturer’s protocol.

    Techniques: In Vitro, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Enzyme-linked Immunosorbent Assay

    GRN aggravated lupus nephritis in lupus model by enhancing ALD-DNA-induced M2b polarization. CD11b + /F4/80 high renal macrophages were sorted from nephritic single-cell suspensions from pGRN-treated, pcDNA3.1-treated, or LV-shGRN-injected, LV-shNC-injected lupus model by flow cytometry. (A) mRNA levels of TNF-α , IL-1β , IL-6 , IL-10 , IL-12 , MCP-1 , Nos2 (iNOS) and Agr1 in the renal macrophages screened from pGRN or pcDNA3.1-treated lupus mice were evaluated by real-time PCR. Data are means ± SD from 8 mice in each group. (B) Real time analysis for the mRNA levels of TNF-α , IL-1β , IL-6 , IL-10 , IL-12 , MCP-1 , Nos2 (iNOS) and Agr1 in renal macrophages from LV-shGRN-injected lupus mice and control mice. Data are means ± SD from 8 mice in each group. *, P

    Journal: PLoS ONE

    Article Title: Granulin Exacerbates Lupus Nephritis via Enhancing Macrophage M2b Polarization

    doi: 10.1371/journal.pone.0065542

    Figure Lengend Snippet: GRN aggravated lupus nephritis in lupus model by enhancing ALD-DNA-induced M2b polarization. CD11b + /F4/80 high renal macrophages were sorted from nephritic single-cell suspensions from pGRN-treated, pcDNA3.1-treated, or LV-shGRN-injected, LV-shNC-injected lupus model by flow cytometry. (A) mRNA levels of TNF-α , IL-1β , IL-6 , IL-10 , IL-12 , MCP-1 , Nos2 (iNOS) and Agr1 in the renal macrophages screened from pGRN or pcDNA3.1-treated lupus mice were evaluated by real-time PCR. Data are means ± SD from 8 mice in each group. (B) Real time analysis for the mRNA levels of TNF-α , IL-1β , IL-6 , IL-10 , IL-12 , MCP-1 , Nos2 (iNOS) and Agr1 in renal macrophages from LV-shGRN-injected lupus mice and control mice. Data are means ± SD from 8 mice in each group. *, P

    Article Snippet: Genomic DNA was subjected to quantitative real-time PCR using a Lightcycler480 and SYBR Green system (Roche Diagnostic Systems) following the manufacturer’s protocol.

    Techniques: Injection, Flow Cytometry, Cytometry, Mouse Assay, Real-time Polymerase Chain Reaction

    Analysis of methylation of p16 and MINT loci in colorectal cell lines. ( A ) For p16 promoter analysis, bisulphite-modified cell line DNA was amplified in separate reactions using primers specific for unmethylated (U) or methylated (M) template (methylation-specific PCR - MSP). Examples of methylated cell lines are T84 and RKO; these show amplification in the methylated reaction only. Presence of PCR product in both unmethylated and methylated reactions of HCT116 is indicative of partial methylation at p16 . LIM1215 is shown as an example of a cell line unmethylated at p16 . ( B ) For analysis of methylation at MINT loci, bisulphite-modified DNA was first PCR amplified using appropriate loci-specific primers, and then the PCR products were digested with restriction enzymes that only cut amplicons generated from the methylated template. MINT2 is shown as a representative MINT locus. Resistance to digestion indicates an unmethylated template. Shown are cell lines with partial methylation at MINT 2, CACO2 and RKO, a fully methylated cell line, COLO205, and an unmethylated cell line LIM1215. Undigested amplicons are shown in the last lane. Molecular weight (MW) marker in A and B is pUC19/ Msp I.

    Journal: British Journal of Cancer

    Article Title: CpG island methylation is a common finding in colorectal cancer cell lines

    doi: 10.1038/sj.bjc.6600699

    Figure Lengend Snippet: Analysis of methylation of p16 and MINT loci in colorectal cell lines. ( A ) For p16 promoter analysis, bisulphite-modified cell line DNA was amplified in separate reactions using primers specific for unmethylated (U) or methylated (M) template (methylation-specific PCR - MSP). Examples of methylated cell lines are T84 and RKO; these show amplification in the methylated reaction only. Presence of PCR product in both unmethylated and methylated reactions of HCT116 is indicative of partial methylation at p16 . LIM1215 is shown as an example of a cell line unmethylated at p16 . ( B ) For analysis of methylation at MINT loci, bisulphite-modified DNA was first PCR amplified using appropriate loci-specific primers, and then the PCR products were digested with restriction enzymes that only cut amplicons generated from the methylated template. MINT2 is shown as a representative MINT locus. Resistance to digestion indicates an unmethylated template. Shown are cell lines with partial methylation at MINT 2, CACO2 and RKO, a fully methylated cell line, COLO205, and an unmethylated cell line LIM1215. Undigested amplicons are shown in the last lane. Molecular weight (MW) marker in A and B is pUC19/ Msp I.

    Article Snippet: Genomic DNA (500 ng) was used in PCR with 0.5 μ M of each of the primers 5′-ATATAAACTTGTGGTAGTTCCAGCTGGT-3′ and 5′-ATCAAAGAATGGTCCTGCACC-3′, 2.5 mM MgCl2 , 100 μ M dNTPs, 1.25 U FastStart TaqDNA polymerase (Roche Diagnostics GmBH, Mannheim, Germany) in the reaction buffer provided by the manufacturer.

    Techniques: Methylation, Modification, Amplification, Polymerase Chain Reaction, Generated, Molecular Weight, Marker

    Analysis of hMLH1 methylation and expression in colorectal cell lines. ( A ) Both the A region (top panel) and C region (bottom panel) of the hMLH1 promoter were amplified from bisulphite-treated DNA. Bst U1 restriction enzyme digest was performed to distinguish amplicons from methylated or unmethylated template; BstU1 will cut only amplicons generated from methylated template. Undigested PCR product is shown in the control lane (-). Lane 1 shows an example of a cell line unmethylated at both the A and C regions, DLD1. Lane 2 shows SW403 that is methylated at the A region, but not C. Lane 3 shows LS411 that is partially methylated at both regions and Lane 4 shows a fully methylated cell line, RKO. Peripheral blood (PB) DNA used as a control is always unmethylated at both the A and C regions. MW market is pUC19/ Msp I. ( B ) Representative Western blot analysis of hMLH1 expression in colorectal cell lines with various combinations of A and C region methylations. SW480 has no methylation at either region and has wild-type hMLH1 . HCT116 also is unmethylated at both regions; however, this cell line has a frameshift mutation in one allele of hMLH1 , leading to a reduction in expression levels. Cell lines exhibiting either full (RKO) or partial (LS411) methylation at both the A and C regions show a loss of hMLH1 protein.

    Journal: British Journal of Cancer

    Article Title: CpG island methylation is a common finding in colorectal cancer cell lines

    doi: 10.1038/sj.bjc.6600699

    Figure Lengend Snippet: Analysis of hMLH1 methylation and expression in colorectal cell lines. ( A ) Both the A region (top panel) and C region (bottom panel) of the hMLH1 promoter were amplified from bisulphite-treated DNA. Bst U1 restriction enzyme digest was performed to distinguish amplicons from methylated or unmethylated template; BstU1 will cut only amplicons generated from methylated template. Undigested PCR product is shown in the control lane (-). Lane 1 shows an example of a cell line unmethylated at both the A and C regions, DLD1. Lane 2 shows SW403 that is methylated at the A region, but not C. Lane 3 shows LS411 that is partially methylated at both regions and Lane 4 shows a fully methylated cell line, RKO. Peripheral blood (PB) DNA used as a control is always unmethylated at both the A and C regions. MW market is pUC19/ Msp I. ( B ) Representative Western blot analysis of hMLH1 expression in colorectal cell lines with various combinations of A and C region methylations. SW480 has no methylation at either region and has wild-type hMLH1 . HCT116 also is unmethylated at both regions; however, this cell line has a frameshift mutation in one allele of hMLH1 , leading to a reduction in expression levels. Cell lines exhibiting either full (RKO) or partial (LS411) methylation at both the A and C regions show a loss of hMLH1 protein.

    Article Snippet: Genomic DNA (500 ng) was used in PCR with 0.5 μ M of each of the primers 5′-ATATAAACTTGTGGTAGTTCCAGCTGGT-3′ and 5′-ATCAAAGAATGGTCCTGCACC-3′, 2.5 mM MgCl2 , 100 μ M dNTPs, 1.25 U FastStart TaqDNA polymerase (Roche Diagnostics GmBH, Mannheim, Germany) in the reaction buffer provided by the manufacturer.

    Techniques: Methylation, Expressing, Amplification, Generated, Polymerase Chain Reaction, Western Blot, Mutagenesis