Total RNA extracted from confirmed T2 transgenic plants was used as the template for synthesizing cDNA. The q-RT PCR analysis was completed in a final reaction volume of 20 μL, which included 10 µL TB Green Premix Ex Taq (Tli RNaseH Plus), 0.4 μL PCR forward primer (10 μM), 0.4 μL PCR reverse primer (10 μM), 0.4 μL ROX Reference Dye (50×), 2 μL cDNA template, and 6.8 μL RNase-free water. The q-RT PCR analysis was performed using the Agilent MX3000P PCR instrument, with the following program: 95 °C for 3 min; 40 cycles of 95 °C for 5 s and 60 °C for 30 s (Table S4 ). This was followed by a melting curve analysis involving the following program: 95 °C for 15 s, 60 °C for 1 min, and 60 °C for 30 s. Relative gene expression values were calculated using the 2−ΔΔCt method, and analysis of plant mutation types by Cc-qPCR [22 (link)]. Three separate biological replicates were performed with each treatment. Variance analysis was performed using GraphPad Prism (8.0).
Mx3000p pcr instrument
Manufactured by Agilent Technologies
Sourced in United States
The Mx3000P PCR instrument is a real-time quantitative PCR (qPCR) system designed for versatile and precise nucleic acid analysis. It features a compact design, a Peltier-based thermal cycling system, and a high-performance optical detection system.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using mx3000p pcr instrument
1
Quantitative RT-PCR Analysis of Transgenic Plants
2
Quantitative EGFR Mutation Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Quantitative mutation detection was carried out using ACCBio’s human EGFR gene mutation quantitative detection kit (Beijing ACCB Biotech Ltd. #180626), according to the manufacturer’s protocol. Quantitative RT-PCR was performed on an Mx3000P PCR instrument (Agilent, Santa Clara, CA, USA) with the following settings: 95 °C for 10 min, followed by 40 cycles at 95 °C for 15 s and 60 °C for 1 min.
3
Comprehensive Oncogene Mutation Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mutation profiles of EGFR, KRAS, NRAS, BRAF, and PIK3CA were detected using the corresponding Human Gene Mutations Detection Kit, Human KRAS Gene Mutations Detection Kit, Human NRAS Gene Mutations Detection Kit, Human BRAF Gene Mutations Detection Kit, and Human PIK3CA Gene Mutations Detection Kit (Real-time Fluorescent PCR, Beijing ACCB Biotech Ltd., Beijing, China), respectively. The tests cover 72 hotspot mutations including 45 in exons 18, 19, 20, and 21 of EGFR; 12 in exons 2 and 3 of KRAS; 9 in exons 2 and 3 of NRAS; 5 in exons 9 and 20 of PIK3CA; BRAF V600E. The quantitative PCR was performed on an Mx3000P PCR instrument (Agilent, CA, USA) with the following settings: 95 °C for 10 min; 40 cycles of 95 °C for 15 s and 60 °C for 1 min. The results were interpreted as per the manufacturer's instructions.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!