The RNAiso Plus reagent (TaKaRa) and PrimeScript RT reagent Kit (TaKaRa) were utilized to extract total RNA and generate cDNA according to the official protocols. Extraction of nuclear and cytoplasmic RNA utilized the Ambion® PARIS™ system according to the kit procedures. The quantitative reverse transcriptase–PCR (qRT–PCR) reactions was performed with the SYBR Green PCR Master Mix (TaKaRa) with ABI7500 Fast Real-Time RCR System (Applied Biosystems, USA). The Primer3 system was utilized to design the gene unique primers which were subsequently synthesized through TsingKe Biological Technology (Guangzhou, China). Every survey was conducted in three times, and GAPDH was utilized to normalize the results as internal reference. The relative quantification method which calculated by 2-ΔΔCt was used in analyzing the expression data.
Abi7500 fast real time rcr system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI7500 Fast Real-Time PCR System is a versatile laboratory instrument designed for quantitative real-time PCR (qPCR) analysis. It provides accurate and reliable gene expression quantification and analysis. The system features fast thermal cycling, precise temperature control, and advanced software for data analysis and visualization.
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Lab products found in correlation
3 protocols using abi7500 fast real time rcr system
1
Quantitative Gene Expression Analysis
2
Quantifying Gene Expression by qRT-PCR
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Total RNA was extracted using the RNAiso Plus reagent (TaKaRa) and PrimeScript RT kit (TaKaRa). Next, RNA was reverse transcribed to generate complementary DNA based on the procedure specified by the manufacturer. We used SYBR Green PCR Premix (TaKaRa) to perform qRT-PCR on the ABI7500 Fast Real-Time RCR System (Applied Biosystems, USA). Next, primers were designed using the PubMed database and synthesized by DynaScience Biotechnology (Guangzhou, China). All experiments were performed in triplicates. The results were normalized using GAPDH as an internal control. Finally, we used the 2-ΔΔCt method for the relative quantification of gene expression.
3
Quantitative Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The RNAiso Plus reagent (TaKaRa) and PrimeScript RT reagent Kit (TaKaRa) were utilized to extract total RNA and generate cDNA according to the o cial protocols. Extraction of nuclear and cytoplasmic RNA utilized the Ambion® PARIS™ system according to the kit procedures. The quantitiative reverse transcriptase-PCR (qRT-PCR) reactions was performed with the SYBR Green PCR Master Mix (TaKaRa) with ABI7500 Fast Real-Time RCR System (Applied Biosystems, USA). The Primer3 system was utilized to design the gene unique primers which were subsequently synthesized through TsingKe Biological Technology (Guangzhou, China). Every survey was conducted in three times, and GAPDH was utilized to normalize the results as internal reference. The relative quanti cation method which calculated by 2 -ΔΔCt was used in analyzing the expression data.
CCK-8 assay 2,000 PCa cells were seeded per well in 96-well plates, and Each well contained medium containing 10% FBS with a total volume of 100 ml. According to the counting kit-8 (CK-04, Dojindo) manufacturer's operating instructions, after the cells were adherent to the plates, the original medium was removed, RPMI1640 diluted CCK-8 reagent (10% v/v) in 100 µl was added to the cells, following 2 hours of incubated. Then, the optical density (OD) at 450 nm was measured by a microplate reader (EXL800, BioTek Instruments).
CCK-8 assay 2,000 PCa cells were seeded per well in 96-well plates, and Each well contained medium containing 10% FBS with a total volume of 100 ml. According to the counting kit-8 (CK-04, Dojindo) manufacturer's operating instructions, after the cells were adherent to the plates, the original medium was removed, RPMI1640 diluted CCK-8 reagent (10% v/v) in 100 µl was added to the cells, following 2 hours of incubated. Then, the optical density (OD) at 450 nm was measured by a microplate reader (EXL800, BioTek Instruments).
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