icycler pcr Search Results


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  • 99
    Bio-Rad icycler iq real time pcr detection system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Iq Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 10027 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler iq real time pcr detection system/product/Bio-Rad
    Average 99 stars, based on 10027 article reviews
    Price from $9.99 to $1999.99
    icycler iq real time pcr detection system - by Bioz Stars, 2020-09
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    91
    Bio-Rad icycler real time pcr detection system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 1824 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler real time pcr detection system/product/Bio-Rad
    Average 91 stars, based on 1824 article reviews
    Price from $9.99 to $1999.99
    icycler real time pcr detection system - by Bioz Stars, 2020-09
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    99
    Bio-Rad icycler iq5 real time pcr detection system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Iq5 Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 783 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler iq5 real time pcr detection system/product/Bio-Rad
    Average 99 stars, based on 783 article reviews
    Price from $9.99 to $1999.99
    icycler iq5 real time pcr detection system - by Bioz Stars, 2020-09
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    91
    Bio-Rad icycler real time pcr machine
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Real Time Pcr Machine, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 711 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler real time pcr machine/product/Bio-Rad
    Average 91 stars, based on 711 article reviews
    Price from $9.99 to $1999.99
    icycler real time pcr machine - by Bioz Stars, 2020-09
    91/100 stars
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    89
    Bio-Rad icycler iq5 multicolor real time pcr detection system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Iq5 Multicolor Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 368 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler iq5 multicolor real time pcr detection system/product/Bio-Rad
    Average 89 stars, based on 368 article reviews
    Price from $9.99 to $1999.99
    icycler iq5 multicolor real time pcr detection system - by Bioz Stars, 2020-09
    89/100 stars
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    89
    Bio-Rad icycler iq5 real time pcr system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Iq5 Real Time Pcr System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 325 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler iq5 real time pcr system/product/Bio-Rad
    Average 89 stars, based on 325 article reviews
    Price from $9.99 to $1999.99
    icycler iq5 real time pcr system - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    89
    Bio-Rad icycler myiq real time pcr detection system
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Myiq Real Time Pcr Detection System, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 311 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler myiq real time pcr detection system/product/Bio-Rad
    Average 89 stars, based on 311 article reviews
    Price from $9.99 to $1999.99
    icycler myiq real time pcr detection system - by Bioz Stars, 2020-09
    89/100 stars
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    88
    Bio-Rad icycler pcr thermocycler
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Pcr Thermocycler, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 88/100, based on 134 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler pcr thermocycler/product/Bio-Rad
    Average 88 stars, based on 134 article reviews
    Price from $9.99 to $1999.99
    icycler pcr thermocycler - by Bioz Stars, 2020-09
    88/100 stars
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    89
    Bio-Rad icycler iq5 pcr thermal cycler
    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well <t>PCR</t> plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an <t>iCycler</t> iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.
    Icycler Iq5 Pcr Thermal Cycler, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 89/100, based on 150 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/icycler iq5 pcr thermal cycler/product/Bio-Rad
    Average 89 stars, based on 150 article reviews
    Price from $9.99 to $1999.99
    icycler iq5 pcr thermal cycler - by Bioz Stars, 2020-09
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    Image Search Results


    Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.

    Journal: Molecular Genetics & Genomic Medicine

    Article Title: Biochemical phenotype of a common disease-causing mutation and a possible therapeutic approach for the phosphomannomutase 2-associated disorder of glycosylation

    doi: 10.1002/mgg3.3

    Figure Lengend Snippet: Ligand binding can affect the thermal stability of PMM2. Heat-induced melting profile of wild-type PMM2 (A and C) and F119L-PMM2 (B and D) were recorded by thermal shift assay and by circular dichroism. For thermal shift assay, the proteins (0.2 mg/mL) were equilibrated in buffer (HEPES 20 mmol/L, NaCl 150 mmol/L, pH 7.5) containing Sypro Orange2.5X and the appropriate ligands: MgCl 2 5 mmol/L, EDTA 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L, Glc-6-P 0.5 mmol/L + MgCl 2 5 mmol/L + vanadate 0.5 mmol/L, Glu-1,6-P 0.5 mmol/L + MgCl 2 5 mmol/L. The samples were distributed in 96-well PCR plates, the plates were sealed, and heated from 25 to 80° at 0.5°C/min. The experiment was run on an iCycler iQ Real Time PCR Detection System. An excitation wavelength of 490 nm and an emission wavelength of 575 nm were used to collect the data. When the melting profile was obtained by circular dichroism the proteins (0.2 mg/mL) were equilibrated in the same buffer in the presence of MgCl 2 1 mmol/L or EDTA 5 mmol/L. The signal at 222 nm was recorded while temperature was increased at 0.5°C/min from 20°C. The raw data were corrected by taking into account the slopes of the pre- and post-transition baselines, then they were normalized.

    Article Snippet: Thermal shift assay was performed by using an iCycler iQ Real Time PCR Detection System (Bio-Rad, Hercules, CA).

    Techniques: Ligand Binding Assay, Thermal Shift Assay, Gas Chromatography, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    Transcriptional analysis of DEV UL55 gene in infected cells in vitro . The transcriptional analysis of DEV UL55 in infected DEFs were performed by qRT-PCR. The transcriptional expression of DEV UL55 gene was normalized to a reference gene(β-actin) and relative to NTC control. The average relative content of the DEV UL55 gene transcripts were calculated at 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12, 16, 24, 36, 48, and 60 h post-infection using iCycler IQ 5.

    Journal: Virology Journal

    Article Title: Establishment of real-time quantitative reverse transcription polymerase chain reaction assay for transcriptional analysis of duck enteritis virus UL55 gene

    doi: 10.1186/1743-422X-8-266

    Figure Lengend Snippet: Transcriptional analysis of DEV UL55 gene in infected cells in vitro . The transcriptional analysis of DEV UL55 in infected DEFs were performed by qRT-PCR. The transcriptional expression of DEV UL55 gene was normalized to a reference gene(β-actin) and relative to NTC control. The average relative content of the DEV UL55 gene transcripts were calculated at 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12, 16, 24, 36, 48, and 60 h post-infection using iCycler IQ 5.

    Article Snippet: The RT-PCR was performed in an iCycler IQ Multicolor Real-Time PCR Detection System(Bio-Rad, USA) with a recommended 20 μl protocol according to the manufacturer's protocol: SYBR Green I Mix 9 μl, each of the primer 20 pM, 1 μl standard template, and finally autoclaved double-filtered nanopure water was added to get the final volume of 20 μl.

    Techniques: Infection, In Vitro, Quantitative RT-PCR, Expressing

    The standard curves of DEV UL55 gene and β-actin by qRT-PCR . A , The standard curve of DEV UL55 gene was calculated by iCycler IQ 5 software. Each dot represents the result of triplicate amplification of each dilution. The standard curve equation is Y = -3.321X + 7.755, the correlation coefficient and the slope value of the regression curve were calculated and indicated. B , The standard curve of reference gene β-actin was calculated by iCycler IQ 5 software. Each dot represents the result of triplicate amplification of each dilution. The standard curve equation is Y = -3.279X+6.465, the correlation coefficient and the slope value of the regression curve were calculated and indicated.

    Journal: Virology Journal

    Article Title: Establishment of real-time quantitative reverse transcription polymerase chain reaction assay for transcriptional analysis of duck enteritis virus UL55 gene

    doi: 10.1186/1743-422X-8-266

    Figure Lengend Snippet: The standard curves of DEV UL55 gene and β-actin by qRT-PCR . A , The standard curve of DEV UL55 gene was calculated by iCycler IQ 5 software. Each dot represents the result of triplicate amplification of each dilution. The standard curve equation is Y = -3.321X + 7.755, the correlation coefficient and the slope value of the regression curve were calculated and indicated. B , The standard curve of reference gene β-actin was calculated by iCycler IQ 5 software. Each dot represents the result of triplicate amplification of each dilution. The standard curve equation is Y = -3.279X+6.465, the correlation coefficient and the slope value of the regression curve were calculated and indicated.

    Article Snippet: The RT-PCR was performed in an iCycler IQ Multicolor Real-Time PCR Detection System(Bio-Rad, USA) with a recommended 20 μl protocol according to the manufacturer's protocol: SYBR Green I Mix 9 μl, each of the primer 20 pM, 1 μl standard template, and finally autoclaved double-filtered nanopure water was added to get the final volume of 20 μl.

    Techniques: Quantitative RT-PCR, Software, Amplification

    Depletion of IL-8 in PC-3 and DU145 cells by siRNA transfection : A and C: Decrease in IL-8 mRNA levels : Cells were transfected with 50 nM of control-siRNA (C-siRNA) or Smartpool siRNA against IL-8 (IL-8 siRNA). Total RNA was extracted 48 h after transfection to determine the levels of IL-8 mRNA by Q-RT PCR, as described in Materials and Methods (text). Each RNA sample was also simultaneously subjected to Q-RT PCR to determine the level of GAPDH mRNA for normalization. The threshold cycle (Ct) of each reaction generated by the iCycler program (Bio-Rad Inc.) was used to determine the relative mRNA levels in the test samples, using the formula: Fold Difference (FD) = 1-(2 ΔCt ), where, ΔCt = (Ct test RNA - Ct GAPDHRNA ). Final value of IL-8 mRNA, relative to that of GAPDH was expressed as: 1/FD × 100 to obtain integer values, in arbitrary units. B and D : Levels of IL-8 protein secreted into the culture medium , 48 h after transfection. IL-8 levels were measured using an ELISA kit. Levels are expressed as % of untreated control. Untreated PC-3 cells secreted ~ 60 ng IL-8/10 6 PC-3 cells/24 h and ~ 10 ng IL-8/10 6 DU145 cells/24 h.

    Journal: Molecular Cancer

    Article Title: Depletion of intrinsic expression of Interleukin-8 in prostate cancer cells causes cell cycle arrest, spontaneous apoptosis and increases the efficacy of chemotherapeutic drugs

    doi: 10.1186/1476-4598-8-57

    Figure Lengend Snippet: Depletion of IL-8 in PC-3 and DU145 cells by siRNA transfection : A and C: Decrease in IL-8 mRNA levels : Cells were transfected with 50 nM of control-siRNA (C-siRNA) or Smartpool siRNA against IL-8 (IL-8 siRNA). Total RNA was extracted 48 h after transfection to determine the levels of IL-8 mRNA by Q-RT PCR, as described in Materials and Methods (text). Each RNA sample was also simultaneously subjected to Q-RT PCR to determine the level of GAPDH mRNA for normalization. The threshold cycle (Ct) of each reaction generated by the iCycler program (Bio-Rad Inc.) was used to determine the relative mRNA levels in the test samples, using the formula: Fold Difference (FD) = 1-(2 ΔCt ), where, ΔCt = (Ct test RNA - Ct GAPDHRNA ). Final value of IL-8 mRNA, relative to that of GAPDH was expressed as: 1/FD × 100 to obtain integer values, in arbitrary units. B and D : Levels of IL-8 protein secreted into the culture medium , 48 h after transfection. IL-8 levels were measured using an ELISA kit. Levels are expressed as % of untreated control. Untreated PC-3 cells secreted ~ 60 ng IL-8/10 6 PC-3 cells/24 h and ~ 10 ng IL-8/10 6 DU145 cells/24 h.

    Article Snippet: Total RNA was isolated using RNAeasy mini kit (Qiagen Inc., Santa Clara, CA) following 48–72 hr transfection and used for reverse transcription real time PCR (qPCR) using the Bio-Rad iCycler iQ real time PCR system (Bio-Rad, Hercules, CA) with the gene specific primers.

    Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Generated, Enzyme-linked Immunosorbent Assay

    Gene regulation by fatty acids. Myotubes were incubated with fatty acids (100 µM) or BSA (40 µM, control) for 24 h and harvested for RNA isolation. Real-time PCR was performed with platinum Taq polymerase and SYBR green on an iCycler PCR

    Journal: Journal of Lipid Research

    Article Title: Metabolic switching of human myotubes is improved by n-3 fatty acids

    doi: 10.1194/jlr.M003319

    Figure Lengend Snippet: Gene regulation by fatty acids. Myotubes were incubated with fatty acids (100 µM) or BSA (40 µM, control) for 24 h and harvested for RNA isolation. Real-time PCR was performed with platinum Taq polymerase and SYBR green on an iCycler PCR

    Article Snippet: Real-time PCR was performed with platinum Taq polymerase (Invitrogen, Breda, The Netherlands) and SYBR green on an iCycler PCR machine (Bio-Rad Laboratories).

    Techniques: Incubation, Isolation, Real-time Polymerase Chain Reaction, SYBR Green Assay, Polymerase Chain Reaction

    Summary of RT–PCR analysis of in vivo transgene copy number. Standard curves were based on quantifications of the ori amplicons, using replicated sets of template composed of CD2F 1 non-transgenic mouse DNA mixed with λgt10- lacZ (see Figure 3 Materials and methods) to generate haploid standards of 1, 2.5, 5, 10, 20, 40 and 50 copies. Cycle thresholds ( C t ) and efficiency parameters were obtained by iCycler software and show a linear correlation with log copy number values ( R 2 = 0.9924). Shown are average C t values for: ori standards (open circles), single-copy annexin V exon 4 (closed triangle, A); R5 rearrangement (closed circle, R5); interpolated copies of Muta™mouse ori (closed diamond, MM) and 18S (closed box, 18S).

    Journal: Mutagenesis

    Article Title: Characterisation of Muta(TM)Mouse ?gt10-lacZ transgene: evidence for in vivo rearrangements

    doi: 10.1093/mutage/geq048

    Figure Lengend Snippet: Summary of RT–PCR analysis of in vivo transgene copy number. Standard curves were based on quantifications of the ori amplicons, using replicated sets of template composed of CD2F 1 non-transgenic mouse DNA mixed with λgt10- lacZ (see Figure 3 Materials and methods) to generate haploid standards of 1, 2.5, 5, 10, 20, 40 and 50 copies. Cycle thresholds ( C t ) and efficiency parameters were obtained by iCycler software and show a linear correlation with log copy number values ( R 2 = 0.9924). Shown are average C t values for: ori standards (open circles), single-copy annexin V exon 4 (closed triangle, A); R5 rearrangement (closed circle, R5); interpolated copies of Muta™mouse ori (closed diamond, MM) and 18S (closed box, 18S).

    Article Snippet: Transgene copy number determinations by RT–PCR Segments of endogenous genes and transgenes of Muta™Mouse DNA and in CD2F1 DNA-transgene mixtures (see standards) were quantified using the iCycler iQ RT–PCR detection system (Bio-Rad Laboratories Inc.).

    Techniques: Reverse Transcription Polymerase Chain Reaction, In Vivo, Transgenic Assay, Software