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.
Pph00150f
Manufactured by Qiagen
Sourced in United States
The PPH00150F is a laboratory equipment item manufactured by Qiagen. It serves as a core component for various scientific applications. The detailed specifications and intended use of this product are not available from the information provided.
Automatically generated - may contain errors
Lab products found in correlation
Iscript cdna synthesis kit, by Bio-Rad (1 mentions)
Maxwell rsc simplyrna cells kit, by Promega (1 mentions)
Sybr green pcr master mix, by Thermo Fisher Scientific (1 mentions)
Quantstudio 3 real time pcr system machine, by Thermo Fisher Scientific (1 mentions)
Quantitect reverse transcriptase kit, by Qiagen (1 mentions)
Direct zol rna miniprep plus kit, by Zymo Research (1 mentions)
Rt2 sybr green, by Qiagen (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Rt2 sybr green mastermix, by Qiagen (1 mentions)
Rt2 first strand kit, by Qiagen (1 mentions)
3 protocols using pph00150f
1
Quantifying Progesterone Receptor Expression
2
Transcriptional Analysis of TNIP1 Haplotypes
3
Epigenetic Enzyme Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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AECs and airway-derived fibroblasts were grown in 6-well plates to 80% confluence, at which point RNA was collected using RNeasy Mini Kits (Qiagen). 500 ng of RNA was used to synthesize cDNA using the RT2 First Strand Kit (Qiagen). cDNA was then combined with 2× RT2 SYBR Green Mastermix (Qiagen) and RNase-free water and distributed onto a manufacturer optimized 384-well Human Epigenetic Chromatin Modification Enzymes Focused Array (PAHS-085E-4, Qiagen) pre-loaded with primers targeting 84 genes encoding epigenetic enzymes and 5 housekeeping genes as per manufacturer’s protocol. A complete list of the genes that were analyzed is available in Table S1 (Additional file 1 ). Additionally, to identify gene expression of CREBBP and EP300, cDNA was combined with 2× RT2 SYBR Green Mastermix, RNase-free water, and primers targeting CREBBP (PPH00324F-200, Qiagen), EP300 (PPH00319A-200, Qiagen), hypoxanthine phosphoribosyltransferase 1 (HPRT1, PPH01018C, Qiagen), ribosomal protein L13a (RPL13A, PPH01020B-200, Qiagen), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, PPH00150F, Qiagen) and loaded onto 384-well reaction plates. Data cleaning and housekeeping gene selection is described in Supplementary Methods (Additional file 2 ). Target gene expression was calculated using the delta Ct method: 2^(CtHousekeeping Gene – CtTarget Gene)*10000.
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