RNA was purified from cells using Maxwell RSC simplyRNA Cells Kit (Promega Corporation, Madison, WI, USA) and cDNA was generated using iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA). qPCR was performed with a cDNA equivalent of 50 ng RNA, 1 µM each of the forward and reverse primers and SYBR Green PCR Master Mix (Applied Biosystems, Carlsbad, CA, USA), using a QuantStudio 3 Real-Time PCR System machine (Applied Biosystems), and primers were used against the following targets: GAPDH (QIAGEN – PPH00150F), PGR (QIAGEN – PPH01007F). CT (threshold cycle) values were determined in triplicate samples by subtracting the target gene CT from the GAPDH CT; 2ΔCT was used to determine the expression of PGR relative to GAPDH.
Quantstudio 3 real time pcr system machine
Manufactured by Thermo Fisher Scientific
Sourced in United States
The QuantStudio 3 Real-Time PCR System is a versatile and compact instrument designed for real-time PCR analysis. It is capable of performing gene expression, genotyping, copy number variation, and other types of real-time PCR experiments. The system features a 96-well block format and supports a wide range of sample volumes and plate types.
Automatically generated - may contain errors
Lab products found in correlation
Iscript cdna synthesis kit, by Bio-Rad (3 mentions)
Maxwell rsc simplyrna cells kit, by Promega (3 mentions)
Sybr green pcr master mix, by Thermo Fisher Scientific (3 mentions)
Simplyrna tissue kit, by Promega (2 mentions)
Pph00150f, by Qiagen (1 mentions)
Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
4 protocols using quantstudio 3 real time pcr system machine
1
Quantifying Progesterone Receptor Expression
2
Quantitative Analysis of Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was extracted and purified from cells or xenografts using Maxwell RSC simplyRNA Cells Kit or simplyRNA Tissue Kit (Promega Corporation). cDNA was generated using iScript cDNA Synthesis Kit (Bio-Rad). qPCR was performed with a cDNA equivalent of 50 ng RNA, 1 mmol/L each of the forward and reverse primers, and SYBR Green PCR Master Mix (Applied Biosystems), using a QuantStudio 3 Real-Time PCR System Machine (Applied Biosystems). All primers were purchased from QIAGEN: GAPDH (PPH00150F) PGR (PPH01007F), CCND1 (PPH00128F), VEGFA (PPH00251C), TFF1 (PPH00998C), PDZK1 (PPH08038E), CDK4 (PPH00118F), CDK12 (PPH05712A), DUSP1 (PPH00406A), FTO (PPH16000B), JUNB (PPH00179A), GREB1 (PPH20761F), MYC (PPH00100B), FOS (PPH00094A), BRD2 (PPH09948A). Ct (threshold cycle) values were determined in triplicate samples by subtracting the target gene Ct from the GAPDH Ct; 2–ΔΔCt was used to determine the expression of selected mRNAs relative to GAPDH.
3
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA was isolated using 1 mL TRIzol Reagent (Invitrogen, Carlsbad, CA) per sample following the TRI Reagent Protocol. Applied Biosystems (Foster City, CA) High-Capacity cDNA Reverse Transcription Kit was used to synthesize cDNA as per manufacturer’s instructions. PCR was performed in duplicate using TaqMan Fast Advanced Master Mix (Applied Biosystems) in 384-well plates, as per instructions, and assayed on a QuantStudio 3 Real-Time PCR System machine (Applied Biosystems). Gene primer information is presented in Supplementary Table S7 . Data were analyzed using the 2−∆∆Ct method with housekeeping gene RPL19 and are presented as relative expression compared to the non-stressed control group.
4
Gene Expression Analysis by qPCR
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!