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  • 99
    Millipore dna extract
    Gel image of PCR products amplified with the three customised multiplex PCR assays and visualised with the QIAxcel system. MPI ( left side ) comprises group/family-specific primers for seven taxa: beetles/thrips, spiders, aphids, earthworms, springtails, dipterans, and lacewings. <t>MPII</t> spiders ( upper right side ) covers two spider families, i.e. lycosids and linyphiids, as well as the genus Pachygnatha . MPII beetles/thrips ( lower right side ) addresses six taxa: the carabid genera Poecilus , Bembidion , Pterostichus, and Harpalus , the ladybeetle Coccinella septempunctata as well as thrips ( Frankliniella , Limothrips ). The shortest and longest fragments within each lane represent the two alignment markers (AM; 15 and 3000 bp) as indicated in the left panel . For amplicon lengths see Fig. 1 and Table 3 . Mixes of <t>DNA</t> templates of targets; approximately 1000 ds copies each in PCR
    Dna Extract, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna extract/product/Millipore
    Average 99 stars, based on 202 article reviews
    Price from $9.99 to $1999.99
    dna extract - by Bioz Stars, 2020-08
    99/100 stars
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    99
    Millipore dna extraction
    Cross-plot of the real-time <t>PCR</t> quantification (logarithmic scale) of duplicate <t>DNA</t> samples (extracted using CTAB method) of P. putida DSM8368 in concentrations in the order of 10 7 , 10 8 , 10 9 CFU ml −1 , as pure culture and in sediment matrix. Error bars indicate one standard deviation of two qPCR runs (each with triplicate qPCR reactions, i.e. n = 6) for each duplicate DNA samples (i.e. n = 12 for each point in the graph). The dotted line shows the theoretical recovery of ndoB genes in sediment samples when inhibiting effects are absent.
    Dna Extraction, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1033 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna extraction/product/Millipore
    Average 99 stars, based on 1033 article reviews
    Price from $9.99 to $1999.99
    dna extraction - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    Millipore dna extraction buffer
    MITA/STING and MRP suppressed HBV replication by inducing innate immune signaling. (A-D) The overexpression plasmid pFlag-MITA/STING or pHA-MRP was co-transfected with HBV plasmid pSM2 into <t>Huh7</t> cells. (A) Intracellular HBV core protein, pIRF3, IRF3, pIκB and IκB protein levels were detected by Western Blot with indicated antibody. The expression of MRP and MITA/STING were detected with anti-HA and anti-Flag antibody, respectively. (B) The levels of Ifnb , (C) pro-inflammatory cytokines Tnfa, Il-6 , chemokine Cxcl2 and (D) ISGs Isg56 , Oas1 , MxA , Cxcl10 mRNAs in MRP or MITA overexpressed with or without pSM2 transfected Huh7 cells were detected by qRT-PCR. (E-I) HepG2 cells were transfected with pHA-MRP and pSM2 together and then treated with PDTC. (E) HBV <t>DNA</t> replicative intermediates and mRNA transcripts were detected by Southern blot and Northern blot, respectively. (F) The HBsAg and (G) HBeAg level in supernatant were measured by ELISA after 5-fold dilution of the supernatant. (H) HBcAg and capsid were detected by Western blot. (I) The supernatant mature virions were immunoprecipitated with anti-HBs antibodies and subjected to HBV core-associated DNA extraction, then quantified by real-time PCR. Significant differences were analyzed using a two-tailed unpaired t -test (*, P
    Dna Extraction Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 157 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna extraction buffer/product/Millipore
    Average 99 stars, based on 157 article reviews
    Price from $9.99 to $1999.99
    dna extraction buffer - by Bioz Stars, 2020-08
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    94
    Millipore dna extraction kit
    Genetic alterations within the <t>DNA</t> sequence of BSDL A. Sequences encoding BSDL were amplified from the RNA extracted from pancreatic tumor SOJ-6 cells using RT-PCR and specific primers. The PCR was performed in a personal Robocycler Gradient 96 (Stratagene). Following PCR, migration on agarose gel was performed to visualize amplified fragments. B. Amplicons were purified and their amino-acid sequences deduced after Sanger sequencing. The BSDL-WT amino-acid sequence corresponds to the 2.2 kb <t>amplicon</t> associated with the BSDL-Mut1 sequence, and the BSDL-Mut2 to that of the 1.8 kb band. Star symbol indicates the identical amino-acids. Only the C-terminal sequences are shown.
    Dna Extraction Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna extraction kit/product/Millipore
    Average 94 stars, based on 185 article reviews
    Price from $9.99 to $1999.99
    dna extraction kit - by Bioz Stars, 2020-08
    94/100 stars
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    Image Search Results


    Gel image of PCR products amplified with the three customised multiplex PCR assays and visualised with the QIAxcel system. MPI ( left side ) comprises group/family-specific primers for seven taxa: beetles/thrips, spiders, aphids, earthworms, springtails, dipterans, and lacewings. MPII spiders ( upper right side ) covers two spider families, i.e. lycosids and linyphiids, as well as the genus Pachygnatha . MPII beetles/thrips ( lower right side ) addresses six taxa: the carabid genera Poecilus , Bembidion , Pterostichus, and Harpalus , the ladybeetle Coccinella septempunctata as well as thrips ( Frankliniella , Limothrips ). The shortest and longest fragments within each lane represent the two alignment markers (AM; 15 and 3000 bp) as indicated in the left panel . For amplicon lengths see Fig. 1 and Table 3 . Mixes of DNA templates of targets; approximately 1000 ds copies each in PCR

    Journal: Journal of Pest Science

    Article Title: Diagnostic PCR assays to unravel food web interactions in cereal crops with focus on biological control of aphids

    doi: 10.1007/s10340-015-0685-8

    Figure Lengend Snippet: Gel image of PCR products amplified with the three customised multiplex PCR assays and visualised with the QIAxcel system. MPI ( left side ) comprises group/family-specific primers for seven taxa: beetles/thrips, spiders, aphids, earthworms, springtails, dipterans, and lacewings. MPII spiders ( upper right side ) covers two spider families, i.e. lycosids and linyphiids, as well as the genus Pachygnatha . MPII beetles/thrips ( lower right side ) addresses six taxa: the carabid genera Poecilus , Bembidion , Pterostichus, and Harpalus , the ladybeetle Coccinella septempunctata as well as thrips ( Frankliniella , Limothrips ). The shortest and longest fragments within each lane represent the two alignment markers (AM; 15 and 3000 bp) as indicated in the left panel . For amplicon lengths see Fig. 1 and Table 3 . Mixes of DNA templates of targets; approximately 1000 ds copies each in PCR

    Article Snippet: The PCR protocol of the MPII beetles/thrips assay differed only slightly from the MPII spiders : 1.5 µl of DNA extract and 30 mM TMAC (Sigma-Aldrich) were used in the total volume of 10 µl (plus PCR-grade water to adjust the volume); thermocycling conditions as described above, but with an annealing temperature of 63.5 °C.

    Techniques: Polymerase Chain Reaction, Amplification, Multiplex Assay

    Cross-plot of the real-time PCR quantification (logarithmic scale) of duplicate DNA samples (extracted using CTAB method) of P. putida DSM8368 in concentrations in the order of 10 7 , 10 8 , 10 9 CFU ml −1 , as pure culture and in sediment matrix. Error bars indicate one standard deviation of two qPCR runs (each with triplicate qPCR reactions, i.e. n = 6) for each duplicate DNA samples (i.e. n = 12 for each point in the graph). The dotted line shows the theoretical recovery of ndoB genes in sediment samples when inhibiting effects are absent.

    Journal: FEMS Microbiology Ecology

    Article Title: Re-evaluation of dioxygenase gene phylogeny for the development and validation of a quantitative assay for environmental aromatic hydrocarbon degraders

    doi: 10.1093/femsec/fiv049

    Figure Lengend Snippet: Cross-plot of the real-time PCR quantification (logarithmic scale) of duplicate DNA samples (extracted using CTAB method) of P. putida DSM8368 in concentrations in the order of 10 7 , 10 8 , 10 9 CFU ml −1 , as pure culture and in sediment matrix. Error bars indicate one standard deviation of two qPCR runs (each with triplicate qPCR reactions, i.e. n = 6) for each duplicate DNA samples (i.e. n = 12 for each point in the graph). The dotted line shows the theoretical recovery of ndoB genes in sediment samples when inhibiting effects are absent.

    Article Snippet: To confirm the presence of the DNA in the sample, the purity of the PCR products and the amplification of the correct size gene fragments, DNA extractions and PCR products were routinely analysed using electrophoresis, run at 100 V for approximately 45 min, using 7 μl of product/extract on 1% agarose gels, containing 0.2 μg ml−1 of ethidium bromide (Sigma-Aldrich) DNA stain, and in 1 x Tris-acetate-EDTA buffer.

    Techniques: Real-time Polymerase Chain Reaction, Standard Deviation

    Identification of a tranducing phage particle, φSaBov LUK , harboring linear phage DNA. (A) A schematic map of linear phage DNA, based on PCR results (see below). Coloring of genes is as in Fig. 1 . (B) Based on genome sequencing results of MNKN and CTH96 transductants, various sets of primer (see above map) were designed and tested to locate a linear form of phage DNA containing a bacteriocin gene cluster and LukD/E genes. PCR was positive with primer pairs p1654/p1655 and p1691/p1694 but not with p1651/p1655 and p1691/pseg, indicating a linear form of phage DNA with left flanking near SAB1654, and right flanking near SAB1694. (C) Southern blot analysis of RF122 chromosomal DNA (C) and phage DNA (P) digested with EcoR I restriction enzyme using a probe specific to the lukE gene (the membrane used in this figure is the same as in Fig. 1 ).

    Journal: Scientific Reports

    Article Title: Phage-mediated horizontal transfer of a Staphylococcus aureus virulence-associated genomic island

    doi: 10.1038/srep09784

    Figure Lengend Snippet: Identification of a tranducing phage particle, φSaBov LUK , harboring linear phage DNA. (A) A schematic map of linear phage DNA, based on PCR results (see below). Coloring of genes is as in Fig. 1 . (B) Based on genome sequencing results of MNKN and CTH96 transductants, various sets of primer (see above map) were designed and tested to locate a linear form of phage DNA containing a bacteriocin gene cluster and LukD/E genes. PCR was positive with primer pairs p1654/p1655 and p1691/p1694 but not with p1651/p1655 and p1691/pseg, indicating a linear form of phage DNA with left flanking near SAB1654, and right flanking near SAB1694. (C) Southern blot analysis of RF122 chromosomal DNA (C) and phage DNA (P) digested with EcoR I restriction enzyme using a probe specific to the lukE gene (the membrane used in this figure is the same as in Fig. 1 ).

    Article Snippet: Phage DNA extraction and PCR The mitomycin C treated culture lysates were treated with excessive amounts of RNase and DNase I (Sigma-Aldrich, 100 unit each), and then phage particles were precipitated with NaCl (0.5 M final concentration) and polyethylene glycol 8000 (10%, wt/vol), followed by ultracentrifugation at 100,000 × g for 1 h. Phage DNA was extracted using DNeasy kit (Qiagen) according to the manufacturers' instructions.

    Techniques: Polymerase Chain Reaction, Sequencing, Southern Blot

    Heterogeneous excision products of the phage (φSaBov) that integrates at genomic island νSaβ. (A) A schematic map of νSaβ in the strain RF122. The arrows represent genes annotated in the GenBank entries 10 and colored based on key features. Orange; restriction modification system HsdR/M, yellow; serine protease cluster ( spl ), light green; bacteriocin gene cluster ( bsa ), pink; leukocidins ( lukD/E ), red; enterotoxin gene cluster ( egc ), cyan; genes related to phage. Direct repeat sequences associated with phage and those associated with the egc were annotated as attN L and attN R and attEGC L and attEGC R , respectively. Sequence variations in the direct repeats were underlined. Primers used for outward PCR and sequencing results of attN P and attEGC p were depicted. (B) Transmission electron microscope analysis of phage particles induced from the strain RF122. At least, three different head sizes (a, b, and c; 58, 47, 26 nm, respectively) of phages were observed. (C) Results of outward PCR using pInt/p1702 and p1693/p1759 for φSaBov N and φSaBov EGC , respectively. (D) RF122 chromosomal DNA (C) and phage DNA (P) were digested with EcoR I, separated by electrophoresis, and transferred to Nylon membrane for Southern blot analysis. Probes specific to the integrase gene (SAB1760, for φSaBov N ), SAB1737 (for φSaBov N and φSaBov EGC ), and the sem gene (for φSaBov EGC ) were used. (E) Phage spot test. Mitomycin C induced culture lysate from the strain RF122 (10 8 pfu/ml) was dropped onto the lawn culture of human ST36-SCCmecII (USA200), ST8-SCCmecIV (USA300), ST1-SCCmecIV (USA400), and bovine mastitis isolate (ST151).

    Journal: Scientific Reports

    Article Title: Phage-mediated horizontal transfer of a Staphylococcus aureus virulence-associated genomic island

    doi: 10.1038/srep09784

    Figure Lengend Snippet: Heterogeneous excision products of the phage (φSaBov) that integrates at genomic island νSaβ. (A) A schematic map of νSaβ in the strain RF122. The arrows represent genes annotated in the GenBank entries 10 and colored based on key features. Orange; restriction modification system HsdR/M, yellow; serine protease cluster ( spl ), light green; bacteriocin gene cluster ( bsa ), pink; leukocidins ( lukD/E ), red; enterotoxin gene cluster ( egc ), cyan; genes related to phage. Direct repeat sequences associated with phage and those associated with the egc were annotated as attN L and attN R and attEGC L and attEGC R , respectively. Sequence variations in the direct repeats were underlined. Primers used for outward PCR and sequencing results of attN P and attEGC p were depicted. (B) Transmission electron microscope analysis of phage particles induced from the strain RF122. At least, three different head sizes (a, b, and c; 58, 47, 26 nm, respectively) of phages were observed. (C) Results of outward PCR using pInt/p1702 and p1693/p1759 for φSaBov N and φSaBov EGC , respectively. (D) RF122 chromosomal DNA (C) and phage DNA (P) were digested with EcoR I, separated by electrophoresis, and transferred to Nylon membrane for Southern blot analysis. Probes specific to the integrase gene (SAB1760, for φSaBov N ), SAB1737 (for φSaBov N and φSaBov EGC ), and the sem gene (for φSaBov EGC ) were used. (E) Phage spot test. Mitomycin C induced culture lysate from the strain RF122 (10 8 pfu/ml) was dropped onto the lawn culture of human ST36-SCCmecII (USA200), ST8-SCCmecIV (USA300), ST1-SCCmecIV (USA400), and bovine mastitis isolate (ST151).

    Article Snippet: Phage DNA extraction and PCR The mitomycin C treated culture lysates were treated with excessive amounts of RNase and DNase I (Sigma-Aldrich, 100 unit each), and then phage particles were precipitated with NaCl (0.5 M final concentration) and polyethylene glycol 8000 (10%, wt/vol), followed by ultracentrifugation at 100,000 × g for 1 h. Phage DNA was extracted using DNeasy kit (Qiagen) according to the manufacturers' instructions.

    Techniques: Modification, Sequencing, Polymerase Chain Reaction, Transmission Assay, Microscopy, Electrophoresis, Southern Blot, Spot Test

    Schematic of small DNA fragment (SDF) generation and PCR analysis of SFHR A. SDF (783bp) containing the ΔF508 mutation and a KpnI restriction enzyme cleavage site was synthesized using primers mCF1 and mCF15, localized within introns 9 and 10

    Journal:

    Article Title: CFTR gene targeting in mouse embryonic stem cells mediated by Small Fragment Homologous Replacement (SFHR)

    doi:

    Figure Lengend Snippet: Schematic of small DNA fragment (SDF) generation and PCR analysis of SFHR A. SDF (783bp) containing the ΔF508 mutation and a KpnI restriction enzyme cleavage site was synthesized using primers mCF1 and mCF15, localized within introns 9 and 10

    Article Snippet: Before transfection the SDF was used, always gel and ethanol purified (DNA gel extraction kit; Millipore, Bedford, MA).

    Techniques: Polymerase Chain Reaction, Mutagenesis, Synthesized

    MITA/STING and MRP suppressed HBV replication by inducing innate immune signaling. (A-D) The overexpression plasmid pFlag-MITA/STING or pHA-MRP was co-transfected with HBV plasmid pSM2 into Huh7 cells. (A) Intracellular HBV core protein, pIRF3, IRF3, pIκB and IκB protein levels were detected by Western Blot with indicated antibody. The expression of MRP and MITA/STING were detected with anti-HA and anti-Flag antibody, respectively. (B) The levels of Ifnb , (C) pro-inflammatory cytokines Tnfa, Il-6 , chemokine Cxcl2 and (D) ISGs Isg56 , Oas1 , MxA , Cxcl10 mRNAs in MRP or MITA overexpressed with or without pSM2 transfected Huh7 cells were detected by qRT-PCR. (E-I) HepG2 cells were transfected with pHA-MRP and pSM2 together and then treated with PDTC. (E) HBV DNA replicative intermediates and mRNA transcripts were detected by Southern blot and Northern blot, respectively. (F) The HBsAg and (G) HBeAg level in supernatant were measured by ELISA after 5-fold dilution of the supernatant. (H) HBcAg and capsid were detected by Western blot. (I) The supernatant mature virions were immunoprecipitated with anti-HBs antibodies and subjected to HBV core-associated DNA extraction, then quantified by real-time PCR. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Journal: PLoS ONE

    Article Title: MITA/STING and Its Alternative Splicing Isoform MRP Restrict Hepatitis B Virus Replication

    doi: 10.1371/journal.pone.0169701

    Figure Lengend Snippet: MITA/STING and MRP suppressed HBV replication by inducing innate immune signaling. (A-D) The overexpression plasmid pFlag-MITA/STING or pHA-MRP was co-transfected with HBV plasmid pSM2 into Huh7 cells. (A) Intracellular HBV core protein, pIRF3, IRF3, pIκB and IκB protein levels were detected by Western Blot with indicated antibody. The expression of MRP and MITA/STING were detected with anti-HA and anti-Flag antibody, respectively. (B) The levels of Ifnb , (C) pro-inflammatory cytokines Tnfa, Il-6 , chemokine Cxcl2 and (D) ISGs Isg56 , Oas1 , MxA , Cxcl10 mRNAs in MRP or MITA overexpressed with or without pSM2 transfected Huh7 cells were detected by qRT-PCR. (E-I) HepG2 cells were transfected with pHA-MRP and pSM2 together and then treated with PDTC. (E) HBV DNA replicative intermediates and mRNA transcripts were detected by Southern blot and Northern blot, respectively. (F) The HBsAg and (G) HBeAg level in supernatant were measured by ELISA after 5-fold dilution of the supernatant. (H) HBcAg and capsid were detected by Western blot. (I) The supernatant mature virions were immunoprecipitated with anti-HBs antibodies and subjected to HBV core-associated DNA extraction, then quantified by real-time PCR. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Article Snippet: Detection of HBV DNA replicative intermediates and HBV core-associated DNA by Southern blot or real-time PCR For extraction of HBV replicative intermediates, Huh7 or HepG2.2.15 cells were lysed in 800 μl of DNA extraction buffer (50 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, 1% NP-40, pH 7.4) with 10 mM MgCl2 and 100 μg/ml DNase I (Sigma-Aldrich, St. Louis, MO) and maintained at 37°C for 0.5 h. The DNase I digestion was stopped using 25 mM EDTA (pH 8.5).

    Techniques: Over Expression, Plasmid Preparation, Transfection, Western Blot, Expressing, Quantitative RT-PCR, Southern Blot, Northern Blot, Enzyme-linked Immunosorbent Assay, Immunoprecipitation, DNA Extraction, Real-time Polymerase Chain Reaction, Two Tailed Test

    MITA/STING, MRP and c-di-GMP inhibited HBV replication in HepG2.2.15 cells. (A-E) The overexpression plasmid pHA-MRP, pFlag-MITA/STING or vector plasmid was transfected into HepG2.2.15 cells. The pHA-β-actin was cotransfected as an inner control. (A) The levels of HBV capsid, intracellular core antigen, pIRF3, IRF3, pIκBα, IκBα, phospho-p65, p65, MRP, MITA, GAPDH and β-actin were detected by immunoblotting with indicated antibodies. (B) The HBV mRNA replicative intermediates were detected by Northern blot. (C) Intracellular HBV core-associated DNA were extracted and quantified with real-time PCR. (D) The HBV mature particles in supernatant were immunoprecipitated with anti-HBs, and then HBV core-associated DNA were extracted and quantified with real-time PCR. (E-G) HepG2.2.15 cells were transfected with c-di-GMP of different concentrations and incubated for 24 h. (E) The HBV DNA replicative intermediates (upper) and mRNAs (lower) were investigated by Southern blot and Northern blot, respectively. (F) HBcAg (upper) and HBV capsid (lower) were immunoblotted with indicated antibody. (G) The supernatant HBV mature particles were immunoprecipitated and HBV core-associated DNA were extracted and quantified with real-time PCR. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Journal: PLoS ONE

    Article Title: MITA/STING and Its Alternative Splicing Isoform MRP Restrict Hepatitis B Virus Replication

    doi: 10.1371/journal.pone.0169701

    Figure Lengend Snippet: MITA/STING, MRP and c-di-GMP inhibited HBV replication in HepG2.2.15 cells. (A-E) The overexpression plasmid pHA-MRP, pFlag-MITA/STING or vector plasmid was transfected into HepG2.2.15 cells. The pHA-β-actin was cotransfected as an inner control. (A) The levels of HBV capsid, intracellular core antigen, pIRF3, IRF3, pIκBα, IκBα, phospho-p65, p65, MRP, MITA, GAPDH and β-actin were detected by immunoblotting with indicated antibodies. (B) The HBV mRNA replicative intermediates were detected by Northern blot. (C) Intracellular HBV core-associated DNA were extracted and quantified with real-time PCR. (D) The HBV mature particles in supernatant were immunoprecipitated with anti-HBs, and then HBV core-associated DNA were extracted and quantified with real-time PCR. (E-G) HepG2.2.15 cells were transfected with c-di-GMP of different concentrations and incubated for 24 h. (E) The HBV DNA replicative intermediates (upper) and mRNAs (lower) were investigated by Southern blot and Northern blot, respectively. (F) HBcAg (upper) and HBV capsid (lower) were immunoblotted with indicated antibody. (G) The supernatant HBV mature particles were immunoprecipitated and HBV core-associated DNA were extracted and quantified with real-time PCR. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Article Snippet: Detection of HBV DNA replicative intermediates and HBV core-associated DNA by Southern blot or real-time PCR For extraction of HBV replicative intermediates, Huh7 or HepG2.2.15 cells were lysed in 800 μl of DNA extraction buffer (50 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, 1% NP-40, pH 7.4) with 10 mM MgCl2 and 100 μg/ml DNase I (Sigma-Aldrich, St. Louis, MO) and maintained at 37°C for 0.5 h. The DNase I digestion was stopped using 25 mM EDTA (pH 8.5).

    Techniques: Over Expression, Plasmid Preparation, Transfection, Northern Blot, Real-time Polymerase Chain Reaction, Immunoprecipitation, Incubation, Southern Blot, Two Tailed Test

    MITA/STING and MRP inhibited HBV replication in vitro . (A-C) The overexpression plasmid pFlag-MITA/STING or pHA-MRP was co-transfected with HBV plasmid pSM2 into Huh7 cells. (A) HBV DNA replicative intermediates (upper) and mRNAs (lower) were detected by Southern blot and Northern blot, respectively. HBV mRNA density signals on Northern blot were normalized to 18S/28S and showed as RNA density ratio. (B) Levels of HBeAg and (C) HBsAg secreted into supernatant were detected by ELISA after 5-fold dilution. Results were presented as the optical density at 450 nm (OD450). (D-G) siRNA specific to human MITA/STING or MRP was transfected into Huh7 for twice. pSM2 was cotransfected with siRNA at the second time. (D) Intracellular HBV core-associated DNA was extracted and quantified with real-time PCR. (E) Intracellular HBV core protein was detected by Western blot. (F) HBsAg and (G) HBeAg expressed in supernatant were diluted 5-fold and measured by ELISA. The dashed line represents the cutoff value (CoV), which was assumed to be 2.1-fold mean value of the negative samples. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Journal: PLoS ONE

    Article Title: MITA/STING and Its Alternative Splicing Isoform MRP Restrict Hepatitis B Virus Replication

    doi: 10.1371/journal.pone.0169701

    Figure Lengend Snippet: MITA/STING and MRP inhibited HBV replication in vitro . (A-C) The overexpression plasmid pFlag-MITA/STING or pHA-MRP was co-transfected with HBV plasmid pSM2 into Huh7 cells. (A) HBV DNA replicative intermediates (upper) and mRNAs (lower) were detected by Southern blot and Northern blot, respectively. HBV mRNA density signals on Northern blot were normalized to 18S/28S and showed as RNA density ratio. (B) Levels of HBeAg and (C) HBsAg secreted into supernatant were detected by ELISA after 5-fold dilution. Results were presented as the optical density at 450 nm (OD450). (D-G) siRNA specific to human MITA/STING or MRP was transfected into Huh7 for twice. pSM2 was cotransfected with siRNA at the second time. (D) Intracellular HBV core-associated DNA was extracted and quantified with real-time PCR. (E) Intracellular HBV core protein was detected by Western blot. (F) HBsAg and (G) HBeAg expressed in supernatant were diluted 5-fold and measured by ELISA. The dashed line represents the cutoff value (CoV), which was assumed to be 2.1-fold mean value of the negative samples. Significant differences were analyzed using a two-tailed unpaired t -test (*, P

    Article Snippet: Detection of HBV DNA replicative intermediates and HBV core-associated DNA by Southern blot or real-time PCR For extraction of HBV replicative intermediates, Huh7 or HepG2.2.15 cells were lysed in 800 μl of DNA extraction buffer (50 mM Tris-HCl, 50 mM NaCl, 1 mM EDTA, 1% NP-40, pH 7.4) with 10 mM MgCl2 and 100 μg/ml DNase I (Sigma-Aldrich, St. Louis, MO) and maintained at 37°C for 0.5 h. The DNase I digestion was stopped using 25 mM EDTA (pH 8.5).

    Techniques: In Vitro, Over Expression, Plasmid Preparation, Transfection, Southern Blot, Northern Blot, Enzyme-linked Immunosorbent Assay, Real-time Polymerase Chain Reaction, Western Blot, Two Tailed Test

    Genetic alterations within the DNA sequence of BSDL A. Sequences encoding BSDL were amplified from the RNA extracted from pancreatic tumor SOJ-6 cells using RT-PCR and specific primers. The PCR was performed in a personal Robocycler Gradient 96 (Stratagene). Following PCR, migration on agarose gel was performed to visualize amplified fragments. B. Amplicons were purified and their amino-acid sequences deduced after Sanger sequencing. The BSDL-WT amino-acid sequence corresponds to the 2.2 kb amplicon associated with the BSDL-Mut1 sequence, and the BSDL-Mut2 to that of the 1.8 kb band. Star symbol indicates the identical amino-acids. Only the C-terminal sequences are shown.

    Journal: Oncotarget

    Article Title: Expression of truncated bile salt-dependent lipase variant in pancreatic pre-neoplastic lesions

    doi: 10.18632/oncotarget.11777

    Figure Lengend Snippet: Genetic alterations within the DNA sequence of BSDL A. Sequences encoding BSDL were amplified from the RNA extracted from pancreatic tumor SOJ-6 cells using RT-PCR and specific primers. The PCR was performed in a personal Robocycler Gradient 96 (Stratagene). Following PCR, migration on agarose gel was performed to visualize amplified fragments. B. Amplicons were purified and their amino-acid sequences deduced after Sanger sequencing. The BSDL-WT amino-acid sequence corresponds to the 2.2 kb amplicon associated with the BSDL-Mut1 sequence, and the BSDL-Mut2 to that of the 1.8 kb band. Star symbol indicates the identical amino-acids. Only the C-terminal sequences are shown.

    Article Snippet: Amplicon purification and sequencing The amplicons obtained were then purified using the DNA extraction kit (Millipore) after electrophoretic migration on agarose gel, according to the protocol recommended by the supplier.

    Techniques: Sequencing, Amplification, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Migration, Agarose Gel Electrophoresis, Purification