rt-primer 5 Millipore Search Results


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  • 99
    Thermo Fisher superscript iii rtase
    Superscript Iii Rtase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 275 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rt pcr
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Rt Pcr, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1285 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mlv rtase
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Mlv Rtase, supplied by Millipore, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rnazol rt
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Rnazol Rt, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 278 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa primescript rt reagent kit
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Primescript Rt Reagent Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 72258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore rt primer
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Rt Primer, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore quantitative rt pcr readymix
    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on <t>RNA</t> level was determined by real-time <t>RT-PCR</t> in H1299 cells incubated with 10 µM of BMVC4 for
    Quantitative Rt Pcr Readymix, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on RNA level was determined by real-time RT-PCR in H1299 cells incubated with 10 µM of BMVC4 for

    Journal: British Journal of Pharmacology

    Article Title: Induction of senescence in cancer cells by the G-quadruplex stabilizer, BMVC4, is independent of its telomerase inhibitory activity

    doi: 10.1111/j.1476-5381.2012.01997.x

    Figure Lengend Snippet: BMVC4 represses c-myc expression through the formation of quadruplex-forming sequence within the QFS2 of the c-myc promoter. (A) Expression of c-myc on RNA level was determined by real-time RT-PCR in H1299 cells incubated with 10 µM of BMVC4 for

    Article Snippet: For RT-PCR analysis, total RNA was isolated using Trizol reagent (Sigma) and reversely transcribed using random hexamers with a cDNA reverse transcription kit (Applied Biosystems).

    Techniques: Expressing, Sequencing, Quantitative RT-PCR, Incubation

    RT–PCR analysis of FAP-1 mRNA expression in cultured colon cancer cells. Patients no. 1, 3, 6, 13, 16, 20, 21, and 24 belonged to the Fas-sensitive group (sensitive to CH11-induced apoptosis; n =8), while the rest belonged to Fas-refractory group (resistant to CH11-induced apoptosis; n =20).

    Journal: British Journal of Cancer

    Article Title: Expression of FAP-1 by human colon adenocarcinoma: implication for resistance against Fas-mediated apoptosis in cancer

    doi: 10.1038/sj.bjc.6602136

    Figure Lengend Snippet: RT–PCR analysis of FAP-1 mRNA expression in cultured colon cancer cells. Patients no. 1, 3, 6, 13, 16, 20, 21, and 24 belonged to the Fas-sensitive group (sensitive to CH11-induced apoptosis; n =8), while the rest belonged to Fas-refractory group (resistant to CH11-induced apoptosis; n =20).

    Article Snippet: RT–PCR detection of FAP-1 RNA was isolated from primarily cultured colon cancer cells by lysis in guanidine thiocyanate (Sigma Chemical Co, St Louis, MO, USA) followed by phenol extraction and ethanol precipitation. cDNA was synthesised from 2 μ g of total isolated RNA in a 20 μ l reaction mixture containing 4 μ l of 5 × RT reaction buffer, 10 U of Rnasin (Promega Corp., Madison, WI, USA), 200 μ deoxynucleotide triphosphate, 40 p oligodeoxythymidylic acid primer, and 20 U of Moloney murine reverse transcriptase (Promega).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Cell Culture

    Role of Klf4 in iNOS expression upon LPS stimulation . (A) RT-PCR for iNOS mRNA demonstrates a significant decrease in iNOS mRNA levels upon Klf4 knockdown compared to LPS-treated cells. (B) Immunoblot showing iNOS levels under different conditions. There is a significant decrease in iNOS proteins levels in Si+LPS cells compared to LPS-treated cells. The graphs represent relative iNOS mRNA and protein levels with respect to the untreated controls. (C) Nitrite assay using Griess reagent was carried out to measure iNOS activity. A significant reduction is noticed in NO production as a result of Klf4 knockdown in LPS-treated samples. (D) Luciferase assay for iNOS promoter activity. Data is represented as relative luciferase units/amount of protein (in μg). A more-than-3-fold decrease is observed in luciferase activity in Si+LPS cells compared to LPS-treated cells. (E) EMSA carried out with nuclear extracts of control and LPS-treated BV-2 cells. Lane 1 shows free iNOS probe, whereas a shift is noticed in lane 2 when nuclear extracts were incubated with the probe. Lane 3 shows a supershift when Klf4-specific antibody was incubated with the probe and the nuclear extracts. The shift and supershift are indicated by arrows. Lane 4 shows a decreased shift when nuclear extracts from unstimulated control BV-2 cells were incubated with the iNOS probe. In lane 5, in addition to biotinylated probe, a 100-molar excess of unbiotinylated (Cold) probe was added along with nuclear extracts from LPS-stimulated cells. No significant band is observed in this lane. *, **, Statistical differences in comparison to control values and #, Statistical differences with respect to LPS-treated values respectively (* p

    Journal: Journal of Neuroinflammation

    Article Title: Kr?ppel-like factor 4, a novel transcription factor regulates microglial activation and subsequent neuroinflammation

    doi: 10.1186/1742-2094-7-68

    Figure Lengend Snippet: Role of Klf4 in iNOS expression upon LPS stimulation . (A) RT-PCR for iNOS mRNA demonstrates a significant decrease in iNOS mRNA levels upon Klf4 knockdown compared to LPS-treated cells. (B) Immunoblot showing iNOS levels under different conditions. There is a significant decrease in iNOS proteins levels in Si+LPS cells compared to LPS-treated cells. The graphs represent relative iNOS mRNA and protein levels with respect to the untreated controls. (C) Nitrite assay using Griess reagent was carried out to measure iNOS activity. A significant reduction is noticed in NO production as a result of Klf4 knockdown in LPS-treated samples. (D) Luciferase assay for iNOS promoter activity. Data is represented as relative luciferase units/amount of protein (in μg). A more-than-3-fold decrease is observed in luciferase activity in Si+LPS cells compared to LPS-treated cells. (E) EMSA carried out with nuclear extracts of control and LPS-treated BV-2 cells. Lane 1 shows free iNOS probe, whereas a shift is noticed in lane 2 when nuclear extracts were incubated with the probe. Lane 3 shows a supershift when Klf4-specific antibody was incubated with the probe and the nuclear extracts. The shift and supershift are indicated by arrows. Lane 4 shows a decreased shift when nuclear extracts from unstimulated control BV-2 cells were incubated with the iNOS probe. In lane 5, in addition to biotinylated probe, a 100-molar excess of unbiotinylated (Cold) probe was added along with nuclear extracts from LPS-stimulated cells. No significant band is observed in this lane. *, **, Statistical differences in comparison to control values and #, Statistical differences with respect to LPS-treated values respectively (* p

    Article Snippet: For performing q(RT)-PCR for Klf4 mRNA, Klf4 primers (forward: 5'-TGC CAG ACC AGA TGC AGT CAC- 3', reverse: 5'-GTA GTG CCT GGT CAG TTC ATC- 3', annealing temperature: 60°C, 35 cycles, amplicon size: 286 bp) were procured from Sigma.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Nitration, Activity Assay, Luciferase, Incubation

    Role of Klf4 in mediating inflammation . Knockdown of Klf4 in BV-2 mouse microglial cells using SiRNA against Klf4 mRNA and subsequent decrease in the expression of pro-inflammatory cytokines. (A) q(RT)-PCR demonstrate a significant decrease in Klf4 mRNA levels in Si+LPS cells compared to LPS alone-treated cells. Cells treated with lipofectamine alone served as controls. The graph represents relative Klf4 mRNA expression values normalized to 18S rRNA internal control. (B) Immunoblot analysis of Klf4 protein isolated from BV-2 cells. Klf4 protein levels were decreased significantly in the Si+LPS group compared to LPS alone and to the Sc+LPS group. The graph represents Klf4 protein levels normalized to β-tubulin. No significant differences were observed in Si-alone and Sc-alone conditions compared to control cells for both mRNA and protein levels. (C-E) Cytokine bead array analysis of pro-inflammatory cytokines upon Klf4 knockdown. There is a more-than-two-fold decrease in TNF-α (C) and MCP-1 (D) levels in Klf4 knockdown samples, and a significant three-fold decrease is noticed in the case of IL-6 (E) . *, **, Statistical differences in comparison to control values (* p

    Journal: Journal of Neuroinflammation

    Article Title: Kr?ppel-like factor 4, a novel transcription factor regulates microglial activation and subsequent neuroinflammation

    doi: 10.1186/1742-2094-7-68

    Figure Lengend Snippet: Role of Klf4 in mediating inflammation . Knockdown of Klf4 in BV-2 mouse microglial cells using SiRNA against Klf4 mRNA and subsequent decrease in the expression of pro-inflammatory cytokines. (A) q(RT)-PCR demonstrate a significant decrease in Klf4 mRNA levels in Si+LPS cells compared to LPS alone-treated cells. Cells treated with lipofectamine alone served as controls. The graph represents relative Klf4 mRNA expression values normalized to 18S rRNA internal control. (B) Immunoblot analysis of Klf4 protein isolated from BV-2 cells. Klf4 protein levels were decreased significantly in the Si+LPS group compared to LPS alone and to the Sc+LPS group. The graph represents Klf4 protein levels normalized to β-tubulin. No significant differences were observed in Si-alone and Sc-alone conditions compared to control cells for both mRNA and protein levels. (C-E) Cytokine bead array analysis of pro-inflammatory cytokines upon Klf4 knockdown. There is a more-than-two-fold decrease in TNF-α (C) and MCP-1 (D) levels in Klf4 knockdown samples, and a significant three-fold decrease is noticed in the case of IL-6 (E) . *, **, Statistical differences in comparison to control values (* p

    Article Snippet: For performing q(RT)-PCR for Klf4 mRNA, Klf4 primers (forward: 5'-TGC CAG ACC AGA TGC AGT CAC- 3', reverse: 5'-GTA GTG CCT GGT CAG TTC ATC- 3', annealing temperature: 60°C, 35 cycles, amplicon size: 286 bp) were procured from Sigma.

    Techniques: Expressing, Polymerase Chain Reaction, Isolation

    Role of Klf4 in Cox-2 expression . (A) Semi-quantitative RT-PCR of Cox-2 mRNA indicates a significant decrease in Si+LPS cells compared to LPS-treated cells (B) Immunoblot for Cox-2 in total protein isolates from BV-2 cells. The protein levels of Cox-2 were also found to be significantly reduced within 12 h of LPS treatment in Si+LPS cells compared to LPS-treated cells. The graphs represent relative Cox-2 mRNA and protein levels with respect to the control samples. (C) Luciferase assay for Cox-2 promoter activity using a pCOX301/pGL2 construct. This construct has the luciferase gene directly regulated by Cox-2 promoter. There is a significant reduction in luciferase activity in the Si+LPS condition compared to the LPS-alone condition, indicating that Klf4 may be involved in regulating Cox-2 promoter activity. *, **, Statistical differences in comparison to control values and #, Statistical differences in comparison to LPS-treated values. (* p

    Journal: Journal of Neuroinflammation

    Article Title: Kr?ppel-like factor 4, a novel transcription factor regulates microglial activation and subsequent neuroinflammation

    doi: 10.1186/1742-2094-7-68

    Figure Lengend Snippet: Role of Klf4 in Cox-2 expression . (A) Semi-quantitative RT-PCR of Cox-2 mRNA indicates a significant decrease in Si+LPS cells compared to LPS-treated cells (B) Immunoblot for Cox-2 in total protein isolates from BV-2 cells. The protein levels of Cox-2 were also found to be significantly reduced within 12 h of LPS treatment in Si+LPS cells compared to LPS-treated cells. The graphs represent relative Cox-2 mRNA and protein levels with respect to the control samples. (C) Luciferase assay for Cox-2 promoter activity using a pCOX301/pGL2 construct. This construct has the luciferase gene directly regulated by Cox-2 promoter. There is a significant reduction in luciferase activity in the Si+LPS condition compared to the LPS-alone condition, indicating that Klf4 may be involved in regulating Cox-2 promoter activity. *, **, Statistical differences in comparison to control values and #, Statistical differences in comparison to LPS-treated values. (* p

    Article Snippet: For performing q(RT)-PCR for Klf4 mRNA, Klf4 primers (forward: 5'-TGC CAG ACC AGA TGC AGT CAC- 3', reverse: 5'-GTA GTG CCT GGT CAG TTC ATC- 3', annealing temperature: 60°C, 35 cycles, amplicon size: 286 bp) were procured from Sigma.

    Techniques: Expressing, Quantitative RT-PCR, Luciferase, Activity Assay, Construct

    Degradation of double-stranded (ds) RNA fragments transfected into mosquito cells. A fragment (807 bp) of viral RNA extracted from C6/36 cells was detected through an RT-PCR at 0 h after transfection (hpt) with dsRNA derived from (+) or (−) 5′3′-UTR RNA. The transfected dsRNA had faded at 3 and 6 h after transfection, suggesting that dsRNAs may have been cleaved, and thus generated undetectable short interfering RNAs.

    Journal: BioMed Research International

    Article Title: Cell Type-Dependent RNA Recombination Frequency in the Japanese Encephalitis Virus

    doi: 10.1155/2014/471323

    Figure Lengend Snippet: Degradation of double-stranded (ds) RNA fragments transfected into mosquito cells. A fragment (807 bp) of viral RNA extracted from C6/36 cells was detected through an RT-PCR at 0 h after transfection (hpt) with dsRNA derived from (+) or (−) 5′3′-UTR RNA. The transfected dsRNA had faded at 3 and 6 h after transfection, suggesting that dsRNAs may have been cleaved, and thus generated undetectable short interfering RNAs.

    Article Snippet: Assessment of RNA Stability by an RT-PCR Sequences derived from (+)5′3′-UTR-II RNA were transfected into cells either treated or untreated with an exoribonuclease inhibitor (3′-phosphoadenosine-5′-phosphate, PAP) (Sigma-Aldrich, St. Louis, MO, USA) and incubated for 5 h. RNA was extracted with the TRIzol reagent (5 PRIME, Gaithersburg, MD, USA), and then DNAse (Promega) was added to delete interference of genomic DNA.

    Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Generated

    Quality, purity and allele-specific expression analyses of extensively washed embryonic control samples. (A) RT-PCR amplifying ACT11 and WOX9 , an embryo-specific gene, from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) using 32 PCR cycles. Genomic DNA was used as positive and water as negative control. (B) RT-PCR amplifying TT10 and AT5G42530 , two seed coat-specific genes ( [37] and our own analysis based on [33] ), from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) and a combined seed coat/endosperm sample using 28 and 32 PCR cycles, respectively. Genomic DNA was used as positive and water as negative control. (C) RT-PCR amplifying FWA, AGL46 and AGL62 , three endosperm-specific genes [16] , [38] , [39] , from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) and a combined seed coat/endosperm sample using 28, 32 and 32 PCR cycles, respectively. Genomic DNA was used as positive and water as negative control. In addition, ACT11 was amplified again including the combined seed coat and endosperm sample. (D) Allele-specific expression analysis of AT1G29660 , AT1G72260 , AT2G47115 , AT5G62210 , AT3G20520 , and AT2G17710 in the 6× washed embryonic cDNA libraries. The analyzed gene and the polymorphism between Col-0 and Ler are indicated in the grey box and the direction of the cross and the stage on top of the panel. The polymorphic nucleotide is displayed in bold and underlined below each chromatogram.

    Journal: PLoS Genetics

    Article Title: Genomic Imprinting in the Arabidopsis Embryo Is Partly Regulated by PRC2

    doi: 10.1371/journal.pgen.1003862

    Figure Lengend Snippet: Quality, purity and allele-specific expression analyses of extensively washed embryonic control samples. (A) RT-PCR amplifying ACT11 and WOX9 , an embryo-specific gene, from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) using 32 PCR cycles. Genomic DNA was used as positive and water as negative control. (B) RT-PCR amplifying TT10 and AT5G42530 , two seed coat-specific genes ( [37] and our own analysis based on [33] ), from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) and a combined seed coat/endosperm sample using 28 and 32 PCR cycles, respectively. Genomic DNA was used as positive and water as negative control. (C) RT-PCR amplifying FWA, AGL46 and AGL62 , three endosperm-specific genes [16] , [38] , [39] , from 1× washed and 6× washed reciprocal, embryonic cDNA libraries (2–4 cell stage) and a combined seed coat/endosperm sample using 28, 32 and 32 PCR cycles, respectively. Genomic DNA was used as positive and water as negative control. In addition, ACT11 was amplified again including the combined seed coat and endosperm sample. (D) Allele-specific expression analysis of AT1G29660 , AT1G72260 , AT2G47115 , AT5G62210 , AT3G20520 , and AT2G17710 in the 6× washed embryonic cDNA libraries. The analyzed gene and the polymorphism between Col-0 and Ler are indicated in the grey box and the direction of the cross and the stage on top of the panel. The polymorphic nucleotide is displayed in bold and underlined below each chromatogram.

    Article Snippet: RT-PCR and Sanger Sequencing RT-PCR was performed on diluted cDNA libraries (4 ng/µl) by doing 28 to 34 cycles (94°C for 15 sec, 58°C for 20 sec, and 72°C for 30 sec) followed by 72°C for 5 min. We used Sigma Taq DNA Polymerase and PCR buffer from Sigma-Aldrich and a final concentration of 2 mM MgCl2 , 0.2 mM dNTPs and 0.2–0.4 mM Primer.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Negative Control, Amplification

    GM-CSF treatment augments CpG-mediated Type 1 IFN production. Balb/c mice were treated with the TLR-9 agonist, CpG DNA (200 μg s.c.). After 18 h, mice were sacrificed and total RNA from spleen and intestine were subjected to real-time RT-PCR analysis.

    Journal:

    Article Title: Granulocyte macrophage colony stimulating factor ameliorates DSS induced experimental colitis

    doi: 10.1002/ibd.20279

    Figure Lengend Snippet: GM-CSF treatment augments CpG-mediated Type 1 IFN production. Balb/c mice were treated with the TLR-9 agonist, CpG DNA (200 μg s.c.). After 18 h, mice were sacrificed and total RNA from spleen and intestine were subjected to real-time RT-PCR analysis.

    Article Snippet: Total RNA was isolated from the proximal colon using Trizol (Invitrogen) and subjected to mRNA purification using Oligotex mRNA mini kit (Qiagen). mRNA was then reverse transcribed with SuperScript II reverse transcriptase (Invitrogen Corp) in the presence of random hexamer primers (Invitrogen Corp). cDNA were used for real time RT-PCR analyses using Jumpstart Taq DNA polymerase (Sigma) and SYBR green nucleic acid stain.

    Techniques: Mouse Assay, Quantitative RT-PCR