steponeplus real time polymerase chain reaction system  (Thermo Fisher)


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    Name:
    StepOnePlus Real Time PCR System
    Description:
    The StepOnePlus Real Time PCR System is a 96 well Real Time PCR instrument perfect for both first time and experienced users The StepOnePlus Real Time PCR System can be setup in a variety of configurations and comes ready to use out of the box with intuitive data analysis and instrument control software Utilizing robust LED based 4 color optical recording the StepOnePlus Real Time PCR System is designed to deliver precise quantitative Real Time PCR results for a variety of genomic research applications We supply a Dell Laptop with the StepOnePlus Real Time PCR System Features of the StepOnePlus Real Time PCR System include • Advanced software and instrumentation for performing a wide array of genomic assays• Sensitive 4 color optical LED recording system• Intuitive and robust software perfect for both first time and advanced users• Simple and flexible instrument set up and usageNew StepOne SoftwareThe StepOne Software included with the StepOnePlus Real Time PCR System runs on the Windows XP operating system and provides instrument control data collection and data analysis capabilities analysis software is also compatible with Windows 7 This latest version includes capabilities to collect melt curve data for High Resolution Melt HRM applications and the option to export in Real Time PCR Data Markup Language RDML for compatibility with MIQE guidelines Rapid and Simple Assay Set UpThis remarkably simple Real Time PCR system is designed with an LCD touchscreen and a user friendly interface that brings the power of genetic studies to researchers new to Real Time PCR Fig 1 The intuitive software and protocol wizards help guide new users through their Real Time PCR experiments The StepOnePlus Real Time PCR System also includes a quick start setup so that you start a run immediately and enter plate information at a later time Sensitive LED Based 4 Color Fluorescence ReadingThe StepOnePlus Real Time PCR System utilizes a long life LED based optical system that can record fluorescence from FAM SYBR Green VIC JOE NED TAMRA and ROX dyes This cost effective 4 color 96 well system delivers precise quantitative Real Time PCR results and saves data from all filters in every run without depending on a computer or plate setup It can discriminate between 2 populations of 5 000 and 10 000 template copies of a TaqMan assay with 99 7 confidence Compatible with Many Genomic Analysis TechniquesPerform a variety of standard and demanding genetic analysis research techniques on one instrument using the user friendly StepOnePlus Real Time PCR System and StepOne Software The StepOnePlus System supports many Real Time PCR applications including the following • SNP Genotyping• Gene Expression Analysis• MicroRNA Expression• Protein Expression• Translocation Analysis• Gene Detection• Viral Load AnalysisThe included StepOne Software supports a variety of analysis methods including the following • Standard Curve absolute quantitation • Relative Standard Curve• Comparative Ct ΔΔCt relative quantitation • Genotyping and Presence Absence• Melt Curve Analysis as a standalone application • High Resolution Melting as a standalone application Versatile Instrument Configuration OptionsThe ultra compact footprint of a StepOnePlus System can be installed in multiple distinct configurations providing unmatched flexibility and convenience that can allow a fit in any laboratory Fig 2 The StepOnePlus Real Time PCR System can be used via a PC with a PC connected to a Local Area Network LAN or as a stand alone instrument PC free Each StepOnePlus Real Time PCR System is factory calibrated for optical and thermal accuracy so simply remarkable Real Time PCR results can be obtained right out of the box Energy Efficiency and Space SavingThe StepOnePlus Real Time PCR System draws 38 less energy to process one sample plate when the instrument is in a heated state compared to the Bio Rad CFX96 Real Time PCR Detection System The StepOnePlus Real Time PCR System also has advanced temperature control through use of VeriFlex Blocks technology and has a footprint nearly 15 smaller than Bio Rad CFX96 which in today s crowded laboratories helps you use your laboratory space even more efficiently You also leave a smaller environmental footprint on the Earth Real Time Data Monitoring Dissemination and StorageThe system measures amplification as it occurs allowing you to monitor the progress of your experiment cycle by cycle either on the machine or remotely Your data is stored on the instrument itself and can be viewed and stored automatically via remote access or transferred via email or a USB flash drive Data can also be conveniently exported as PowerPoint Excel and graphical image files Expanded Gene Expression CapabilitiesThe advanced software provided with the StepOnePlus Real Time PCR System now includes the powerful Gene Expression Study Package This software package allows for greater flexibility and accuracy in your gene expression assays through • Analysis of an unlimited number of plates in one study• Sorting of data by biological or technical replicate group• Use of multiple endogenous controls• Assay efficiency correctionAdvanced High Resolution Melting CapabilityNow with High Resolution Melt Software v3 0 you can perform post PCR sequence variation analysis with the StepOnePlus Real Time PCR System This separately purchased software simplifies set up by maintaining assay specific settings and accepts pasted plate layout information directly from Excel HRM Software v3 0 also has an improved clustering algorithm for increased sensitivity and accuracy and the ability to conduct separate analyses of multiple assays run on a single plate
    Catalog Number:
    4376598
    Price:
    None
    Applications:
    Genotyping & Genomic Profiling|PCR & Real-Time PCR|Real Time PCR (qPCR)|Real Time PCR-Based Gene Expression Profiling|Real-Time PCR Instruments, Software & Calibration|Gene Expression Analysis & Genotyping|Genotyping Instruments, Software & Calibration|High Resolution Melting (HRM) Analysis
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    Instruments and Equipment
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    Structured Review

    Thermo Fisher steponeplus real time polymerase chain reaction system
    The StepOnePlus Real Time PCR System is a 96 well Real Time PCR instrument perfect for both first time and experienced users The StepOnePlus Real Time PCR System can be setup in a variety of configurations and comes ready to use out of the box with intuitive data analysis and instrument control software Utilizing robust LED based 4 color optical recording the StepOnePlus Real Time PCR System is designed to deliver precise quantitative Real Time PCR results for a variety of genomic research applications We supply a Dell Laptop with the StepOnePlus Real Time PCR System Features of the StepOnePlus Real Time PCR System include • Advanced software and instrumentation for performing a wide array of genomic assays• Sensitive 4 color optical LED recording system• Intuitive and robust software perfect for both first time and advanced users• Simple and flexible instrument set up and usageNew StepOne SoftwareThe StepOne Software included with the StepOnePlus Real Time PCR System runs on the Windows XP operating system and provides instrument control data collection and data analysis capabilities analysis software is also compatible with Windows 7 This latest version includes capabilities to collect melt curve data for High Resolution Melt HRM applications and the option to export in Real Time PCR Data Markup Language RDML for compatibility with MIQE guidelines Rapid and Simple Assay Set UpThis remarkably simple Real Time PCR system is designed with an LCD touchscreen and a user friendly interface that brings the power of genetic studies to researchers new to Real Time PCR Fig 1 The intuitive software and protocol wizards help guide new users through their Real Time PCR experiments The StepOnePlus Real Time PCR System also includes a quick start setup so that you start a run immediately and enter plate information at a later time Sensitive LED Based 4 Color Fluorescence ReadingThe StepOnePlus Real Time PCR System utilizes a long life LED based optical system that can record fluorescence from FAM SYBR Green VIC JOE NED TAMRA and ROX dyes This cost effective 4 color 96 well system delivers precise quantitative Real Time PCR results and saves data from all filters in every run without depending on a computer or plate setup It can discriminate between 2 populations of 5 000 and 10 000 template copies of a TaqMan assay with 99 7 confidence Compatible with Many Genomic Analysis TechniquesPerform a variety of standard and demanding genetic analysis research techniques on one instrument using the user friendly StepOnePlus Real Time PCR System and StepOne Software The StepOnePlus System supports many Real Time PCR applications including the following • SNP Genotyping• Gene Expression Analysis• MicroRNA Expression• Protein Expression• Translocation Analysis• Gene Detection• Viral Load AnalysisThe included StepOne Software supports a variety of analysis methods including the following • Standard Curve absolute quantitation • Relative Standard Curve• Comparative Ct ΔΔCt relative quantitation • Genotyping and Presence Absence• Melt Curve Analysis as a standalone application • High Resolution Melting as a standalone application Versatile Instrument Configuration OptionsThe ultra compact footprint of a StepOnePlus System can be installed in multiple distinct configurations providing unmatched flexibility and convenience that can allow a fit in any laboratory Fig 2 The StepOnePlus Real Time PCR System can be used via a PC with a PC connected to a Local Area Network LAN or as a stand alone instrument PC free Each StepOnePlus Real Time PCR System is factory calibrated for optical and thermal accuracy so simply remarkable Real Time PCR results can be obtained right out of the box Energy Efficiency and Space SavingThe StepOnePlus Real Time PCR System draws 38 less energy to process one sample plate when the instrument is in a heated state compared to the Bio Rad CFX96 Real Time PCR Detection System The StepOnePlus Real Time PCR System also has advanced temperature control through use of VeriFlex Blocks technology and has a footprint nearly 15 smaller than Bio Rad CFX96 which in today s crowded laboratories helps you use your laboratory space even more efficiently You also leave a smaller environmental footprint on the Earth Real Time Data Monitoring Dissemination and StorageThe system measures amplification as it occurs allowing you to monitor the progress of your experiment cycle by cycle either on the machine or remotely Your data is stored on the instrument itself and can be viewed and stored automatically via remote access or transferred via email or a USB flash drive Data can also be conveniently exported as PowerPoint Excel and graphical image files Expanded Gene Expression CapabilitiesThe advanced software provided with the StepOnePlus Real Time PCR System now includes the powerful Gene Expression Study Package This software package allows for greater flexibility and accuracy in your gene expression assays through • Analysis of an unlimited number of plates in one study• Sorting of data by biological or technical replicate group• Use of multiple endogenous controls• Assay efficiency correctionAdvanced High Resolution Melting CapabilityNow with High Resolution Melt Software v3 0 you can perform post PCR sequence variation analysis with the StepOnePlus Real Time PCR System This separately purchased software simplifies set up by maintaining assay specific settings and accepts pasted plate layout information directly from Excel HRM Software v3 0 also has an improved clustering algorithm for increased sensitivity and accuracy and the ability to conduct separate analyses of multiple assays run on a single plate
    https://www.bioz.com/result/steponeplus real time polymerase chain reaction system/product/Thermo Fisher
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    steponeplus real time polymerase chain reaction system - by Bioz Stars, 2021-01
    99/100 stars

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    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Effects of cinnamoyloxy-mammeisin from geopropolis on osteoclast differentiation and Porphyromonas gingivalis-induced periodontitis
    Article Snippet: .. Quantitative real-time PCR (qPCR) was performed on StepOnePlus™ system using a predesigned TaqMan® primers/probes set (ThermoScientific-Applied Biosystems). .. The nuclear factor of activated T cells, calcineurin-dependent 1 (Nfatc1), tartrate-resistant acid phosphatase (Trap/Acp5), ATPase, H+ transporting, lysosomal V0 subunit D2 (V-ATPase), Ctsk, Integrin beta 3 (Itgb3) and matrix metallopeptidase 9 (MMP-9) was evaluated using glyceraldehyde 3-phosphate dehydrogenase (Gapdh) as an endogenous control (reference gene).

    Article Title: Contribution of histone N-terminal tails to the structure and stability of nucleosomes
    Article Snippet: .. The sample temperature was increased by the StepOnePlus™ Real-Time PCR unit (Applied Biosystems), and the fluorescence signals were measured with this system. .. Since the wavelength at the fluorescence emission maximum of SYPRO Orange is 570 nm, the fluorescence filter ‘filter 3’, which covers the emission wavelength ranges of the TAMRA (580 nm) and NED (575 nm) dyes, was used for detecting the fluorescence of SYPRO Orange bound to the denatured histones.

    Article Title: An EBV recombinant deleted for residues 130-159 in EBNA3C can deregulate p53/Mdm2 and Cyclin D1/CDK6 which results in apoptosis and reduced cell proliferation
    Article Snippet: .. RT-qPCR was performed on a StepOnePlus real-time PCR System (Applied Biosystems Inc, Carlsbad, CA). ..

    Article Title: Reciprocal Inflammatory Signaling Between Intestinal Epithelial Cells and Adipocytes in the Absence of Immune Cells
    Article Snippet: .. Reverse transcription was then performed using High-Capacity cDNA Reverse Transcription Kits (Applied Biosystems), and mRNA levels were quantified by fluorescence real-time PCR on a StepOnePlus (Applied Biosystems) using TaqMan Gene Expression Assays (Applied Biosystems) or PrimeTime qPCR Assays (Integrated DNA Technologies). ..

    Article Title: Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses
    Article Snippet: .. We used the TaqMan® Fast Virus 1-Step master mix (Applied Biosystems) for RT-qPCR amplification with the recommended cycling parameters, 50ºC for 5 min for reverse transcription, 95ºC for 20 s for RT inactivation/initial denaturation, followed by 45 cycles of 95ºC for 3 s and 60ºC for 30 s. RNA (2 μL) was used as a template in a 20 µL reaction, and all assays were performed using the StepOnePlus Real-Time PCR System (Applied Biosystems). .. The assay amplification efficiency was calculated by the standard curve method ( ) using a 10-fold, 8-log, serial dilution starting at 2 × 108 RNA copies/µL, in duplicate.

    Article Title: VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity
    Article Snippet: .. Quantitative real-time PCR The mRNA levels of endogenous genes were detected by quantitative real-time PCR using a StepOnePlus Real-Time PCR System (Applied Biosystems) with the FastStart Universal SYBR Green Master (Roche Applied Science). .. A 20 μl of reaction cocktail was constituted of 3 μl diluted cDNA, 10 μl 2X SYBR Green Master Mix and 0.5 μl each of the forward and reverse primers.

    Article Title: Metabolic Engineering of Escherichia coli for Enhanced Production of Naringenin 7-Sulfate and Its Biological Activities
    Article Snippet: .. Power SYBR Green Master Mix (Thermo Fisher Scientific, United States) was performed by quantitative real-time PCR (qRT-PCR) on StepOnePlus real-time PCR system (Applied Biosystems, United States). ..

    Article Title: Monitoring Antigen-Specific T Cell Responses Using Real-Time PCR
    Article Snippet: .. The following cycling parameters for PCR amplification are utilized on the StepOnePlus™ Real-Time PCR System (Life Technologies) (with melting curve analysis immediately following in order to confirm the absence of nonspecific amplification): 20 s at 95 °C. .. Using specific software from the qRT-PCR system, assign a baseline range that demonstrates little to no fluorescent activity up to the cycle before amplification becomes evident (usually cycles 3–15).

    Amplification:

    Article Title: Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses
    Article Snippet: .. We used the TaqMan® Fast Virus 1-Step master mix (Applied Biosystems) for RT-qPCR amplification with the recommended cycling parameters, 50ºC for 5 min for reverse transcription, 95ºC for 20 s for RT inactivation/initial denaturation, followed by 45 cycles of 95ºC for 3 s and 60ºC for 30 s. RNA (2 μL) was used as a template in a 20 µL reaction, and all assays were performed using the StepOnePlus Real-Time PCR System (Applied Biosystems). .. The assay amplification efficiency was calculated by the standard curve method ( ) using a 10-fold, 8-log, serial dilution starting at 2 × 108 RNA copies/µL, in duplicate.

    Article Title: Monitoring Antigen-Specific T Cell Responses Using Real-Time PCR
    Article Snippet: .. The following cycling parameters for PCR amplification are utilized on the StepOnePlus™ Real-Time PCR System (Life Technologies) (with melting curve analysis immediately following in order to confirm the absence of nonspecific amplification): 20 s at 95 °C. .. Using specific software from the qRT-PCR system, assign a baseline range that demonstrates little to no fluorescent activity up to the cycle before amplification becomes evident (usually cycles 3–15).

    Fluorescence:

    Article Title: Contribution of histone N-terminal tails to the structure and stability of nucleosomes
    Article Snippet: .. The sample temperature was increased by the StepOnePlus™ Real-Time PCR unit (Applied Biosystems), and the fluorescence signals were measured with this system. .. Since the wavelength at the fluorescence emission maximum of SYPRO Orange is 570 nm, the fluorescence filter ‘filter 3’, which covers the emission wavelength ranges of the TAMRA (580 nm) and NED (575 nm) dyes, was used for detecting the fluorescence of SYPRO Orange bound to the denatured histones.

    Article Title: Reciprocal Inflammatory Signaling Between Intestinal Epithelial Cells and Adipocytes in the Absence of Immune Cells
    Article Snippet: .. Reverse transcription was then performed using High-Capacity cDNA Reverse Transcription Kits (Applied Biosystems), and mRNA levels were quantified by fluorescence real-time PCR on a StepOnePlus (Applied Biosystems) using TaqMan Gene Expression Assays (Applied Biosystems) or PrimeTime qPCR Assays (Integrated DNA Technologies). ..

    Quantitative RT-PCR:

    Article Title: An EBV recombinant deleted for residues 130-159 in EBNA3C can deregulate p53/Mdm2 and Cyclin D1/CDK6 which results in apoptosis and reduced cell proliferation
    Article Snippet: .. RT-qPCR was performed on a StepOnePlus real-time PCR System (Applied Biosystems Inc, Carlsbad, CA). ..

    Article Title: Multiplexed reverse transcription real-time polymerase chain reaction for simultaneous detection of Mayaro, Oropouche, and Oropouche-like viruses
    Article Snippet: .. We used the TaqMan® Fast Virus 1-Step master mix (Applied Biosystems) for RT-qPCR amplification with the recommended cycling parameters, 50ºC for 5 min for reverse transcription, 95ºC for 20 s for RT inactivation/initial denaturation, followed by 45 cycles of 95ºC for 3 s and 60ºC for 30 s. RNA (2 μL) was used as a template in a 20 µL reaction, and all assays were performed using the StepOnePlus Real-Time PCR System (Applied Biosystems). .. The assay amplification efficiency was calculated by the standard curve method ( ) using a 10-fold, 8-log, serial dilution starting at 2 × 108 RNA copies/µL, in duplicate.

    Article Title: Metabolic Engineering of Escherichia coli for Enhanced Production of Naringenin 7-Sulfate and Its Biological Activities
    Article Snippet: .. Power SYBR Green Master Mix (Thermo Fisher Scientific, United States) was performed by quantitative real-time PCR (qRT-PCR) on StepOnePlus real-time PCR system (Applied Biosystems, United States). ..

    SYBR Green Assay:

    Article Title: VRK3-mediated nuclear localization of HSP70 prevents glutamate excitotoxicity-induced apoptosis and Aβ accumulation via enhancement of ERK phosphatase VHR activity
    Article Snippet: .. Quantitative real-time PCR The mRNA levels of endogenous genes were detected by quantitative real-time PCR using a StepOnePlus Real-Time PCR System (Applied Biosystems) with the FastStart Universal SYBR Green Master (Roche Applied Science). .. A 20 μl of reaction cocktail was constituted of 3 μl diluted cDNA, 10 μl 2X SYBR Green Master Mix and 0.5 μl each of the forward and reverse primers.

    Article Title: Metabolic Engineering of Escherichia coli for Enhanced Production of Naringenin 7-Sulfate and Its Biological Activities
    Article Snippet: .. Power SYBR Green Master Mix (Thermo Fisher Scientific, United States) was performed by quantitative real-time PCR (qRT-PCR) on StepOnePlus real-time PCR system (Applied Biosystems, United States). ..

    Expressing:

    Article Title: Reciprocal Inflammatory Signaling Between Intestinal Epithelial Cells and Adipocytes in the Absence of Immune Cells
    Article Snippet: .. Reverse transcription was then performed using High-Capacity cDNA Reverse Transcription Kits (Applied Biosystems), and mRNA levels were quantified by fluorescence real-time PCR on a StepOnePlus (Applied Biosystems) using TaqMan Gene Expression Assays (Applied Biosystems) or PrimeTime qPCR Assays (Integrated DNA Technologies). ..

    Polymerase Chain Reaction:

    Article Title: Monitoring Antigen-Specific T Cell Responses Using Real-Time PCR
    Article Snippet: .. The following cycling parameters for PCR amplification are utilized on the StepOnePlus™ Real-Time PCR System (Life Technologies) (with melting curve analysis immediately following in order to confirm the absence of nonspecific amplification): 20 s at 95 °C. .. Using specific software from the qRT-PCR system, assign a baseline range that demonstrates little to no fluorescent activity up to the cycle before amplification becomes evident (usually cycles 3–15).

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  • 99
    Thermo Fisher cdna
    IGSF3 regulation of sphingolipid metabolism. ( A and B ) Heatmap of expression levels of genes involved in sphingolipid ( A ) and ganglioside ( B ) metabolism relative to endogenous IGSF3 expression levels, measured by <t>cDNA</t> microarray in lymphoblastoids from patient and 5 healthy controls. ( C–G ) mRNA expression levels of indicated enzymes determined by targeted <t>qPCR.</t> Dots represent independent experiments; mean ± SEM shown as horizontal lines; * P
    Cdna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10468 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cdna/product/Thermo Fisher
    Average 99 stars, based on 10468 article reviews
    Price from $9.99 to $1999.99
    cdna - by Bioz Stars, 2021-01
    99/100 stars
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    99
    Thermo Fisher real time pcr system
    Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time <t>PCR</t> and <t>TaqMan</t> ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.
    Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10718 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 10718 article reviews
    Price from $9.99 to $1999.99
    real time pcr system - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

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    IGSF3 regulation of sphingolipid metabolism. ( A and B ) Heatmap of expression levels of genes involved in sphingolipid ( A ) and ganglioside ( B ) metabolism relative to endogenous IGSF3 expression levels, measured by cDNA microarray in lymphoblastoids from patient and 5 healthy controls. ( C–G ) mRNA expression levels of indicated enzymes determined by targeted qPCR. Dots represent independent experiments; mean ± SEM shown as horizontal lines; * P

    Journal: JCI Insight

    Article Title: IGSF3 mutation identified in patient with severe COPD alters cell function and motility

    doi: 10.1172/jci.insight.138101

    Figure Lengend Snippet: IGSF3 regulation of sphingolipid metabolism. ( A and B ) Heatmap of expression levels of genes involved in sphingolipid ( A ) and ganglioside ( B ) metabolism relative to endogenous IGSF3 expression levels, measured by cDNA microarray in lymphoblastoids from patient and 5 healthy controls. ( C–G ) mRNA expression levels of indicated enzymes determined by targeted qPCR. Dots represent independent experiments; mean ± SEM shown as horizontal lines; * P

    Article Snippet: A total of 2 μg of total RNA (RNeasy Mini Plus, QIAGEN) was used to reverse transcribe cDNA (High-Capacity cDNA Reverse Transcription, Thermo Fisher Scientific). qPCR was then performed on the cDNA (StepOnePlus System, Thermo Fisher Scientific) using TaqMan Universal PCR Master Mix (Thermo Fisher Scientific) and probes (all from Thermo Fisher Scientific) specific for human IGSF3 (Hs00155437_m1, lot no. 1163927), mouse IGSF3 (Mm01302155_m1, lot no. 1342279), human CERT (Hs01062552_m1, lot no. 156453), human FAPP2 (Hs01696164_s1, lot no. P161121-011 A12), human ITGB1 (Hs00559595_m1), and human GBA (Hs00164683_m1, lot no. 1611503).

    Techniques: Expressing, Microarray, Real-time Polymerase Chain Reaction

    Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time PCR and TaqMan ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.

    Journal: Molecular Medicine Reports

    Article Title: A novel method to detect the Mexican founder mutation BRCA1 ex9-12del associated with breast and ovarian cancer using quantitative polymerase chain reaction and TaqMan® probes

    doi: 10.3892/mmr.2018.9141

    Figure Lengend Snippet: Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time PCR and TaqMan ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.

    Article Snippet: qPCR with designed primers and TaqMan® probes Thermocycler StepOnePlus™ Real-Time PCR System (Thermo Fisher Scientific, Inc.) was used to perform the qPCR with TaqMan® probes.

    Techniques: Genotyping Assay, Mutagenesis, Real-time Polymerase Chain Reaction, Fluorescence, Software, Polymerase Chain Reaction

    Detection of the BRCA1 ex9-12del mutation by real-time PCR and TaqMan ® probe. Red amplification curves represent the fluorescent signal given by the FAM ™ dye from the TaqMan ® probe detecting the variant allele in the four positive samples, which had a mean Ct of 27.46. No amplification curve is observed for the negative samples neither for negative controls (horizontal red lines). The ‘Y’ axis values represent the fluorescent signal normalized of the FAM ™ dye with the ROX, whereas the ‘X’ axis represent number of cycles. PCR. PCR, polymerase chain reaction; ROX, passive reference dye.

    Journal: Molecular Medicine Reports

    Article Title: A novel method to detect the Mexican founder mutation BRCA1 ex9-12del associated with breast and ovarian cancer using quantitative polymerase chain reaction and TaqMan® probes

    doi: 10.3892/mmr.2018.9141

    Figure Lengend Snippet: Detection of the BRCA1 ex9-12del mutation by real-time PCR and TaqMan ® probe. Red amplification curves represent the fluorescent signal given by the FAM ™ dye from the TaqMan ® probe detecting the variant allele in the four positive samples, which had a mean Ct of 27.46. No amplification curve is observed for the negative samples neither for negative controls (horizontal red lines). The ‘Y’ axis values represent the fluorescent signal normalized of the FAM ™ dye with the ROX, whereas the ‘X’ axis represent number of cycles. PCR. PCR, polymerase chain reaction; ROX, passive reference dye.

    Article Snippet: qPCR with designed primers and TaqMan® probes Thermocycler StepOnePlus™ Real-Time PCR System (Thermo Fisher Scientific, Inc.) was used to perform the qPCR with TaqMan® probes.

    Techniques: Mutagenesis, Real-time Polymerase Chain Reaction, Amplification, Variant Assay, Polymerase Chain Reaction

    p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Azacitidine mitigates GvHD via differential effects on the proliferation of T effectors and nTregs in vivo

    doi: 10.4049/jimmunol.1502399

    Figure Lengend Snippet: p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p

    Article Snippet: QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System (Thermo fisher) using pre-designed TaqMan® Gene Expression Assays (Life Technologies) (18S RNA Mm03928990 and p21 Mm04205640) according to manufacturer's instructions.

    Techniques: Isolation, Cell Culture, Recombinant, FACS, RNA Extraction, Real-time Polymerase Chain Reaction, Expressing