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DNA Sequencing confirmed Cd 26 mutation on 5 ambiguous samples and revealed two homozygous mutation.
Bio rad cfx manager gene study software#
When an instrument is connected to the computer, CFX Manager software will check for. The same gene expression analysis choices that are available in the Data Analysis window are available here. Specify the default settings for a new Gene Expression data file. quantification by CFX Manager software 2.1 (Bio-Rad Laboratories). 3t thce e Sl e Study Analysis tab to analyze data. Table 3 shows gene-specific primer sets used (Bio-Rad Laboratories) -actin gene was. In the Study Setup tab (Figure 7), click Add Data Files and browse to select files for inclusion. High resolution melt v3.1 software and Bio-Rad CFX Manager software were used to analyze the result of HRMA and Tm-shift qPCR, whereas the T-ARMS-PCR result was analyzed by observing the number and size of DNA bands on gel electrophoresis.Īmong 21 samples, the Cd 26 mutation was detected in numbers 18, 19 and 21 by HRMA, Tm-shift qPCR and T-ARMS-PCR. Select File > New > Gene Study from the main window.
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Bio rad cfx manager gene study archive#
This study was conducted to compare the detection result of Cd 26 (G→A) mutation in the β-globin-coding gene of heterozygous HbE between the above-mentioned methods.ĭNA samples were isolated from blood archive of heterozygous HbE and analyzed for the detection of the mutation using HRMA and Tm-shift on a real-time PCR instrument, whereas T-ARMS analysis was performed on a conventional PCR equipment. Various types of molecular analysis methods such as tetra-primer amplification refractory mutation system (T-ARMS-PCR), Tm-shift real-time polymerase chain reaction (Tm-shift qPCR) and high-resolution melting analysis (HRMA) are commonly used to detect several mutations in the β-globin-coding gene. BioRad CFX96 Real Time PCR Thermal Cycler quantity. The system is composed of the CFX96 Optical Reaction Module, the C1000 Touch Thermal Cycler Chassis, SYBR®Green Supermix, 96-well PCR plates, and plate seals. Hemoglobin E is a variant hemoglobin caused due to the base substitution G→A at codon 26 in the β-globin-coding gene that is followed by the alteration of glutamic acid (GAG) to lysine (AAG). Bio-Rad CFX96 Touch Real-Time PCR Detection System is used for precise, reproducible real-time PCR.