Total RNA was extracted from tissue samples using the RNeasy minikit (Qiagen, Valencia, CA, USA) and homogenized using a Polytron (Kynematica Polytron, Westbury, USA). Total RNA (0.5 µg) was retro-transcribed (RT) using TaqMan reverse transcription reagents from Applied Biosystems (Branchburg, USA) plus random hexamers. Real-time PCR was performed using the ROCHE sequence detection system with the TaqMan universal PCR master mix and probes (Applied Biosystems) for the mouse Slc27a1/FATP1, Oxct1 and Pdk4 genes, using Rn18s as the endogenous control to normalize the crossing point (CP). For the Hmgcs2 gene real-time PCR was performed using the ABI PRISM 7500 sequence detection system (Applied Biosystems) with the GoTaq qPCR master mix (Promega) and SyberGreen labelling and using the Actb gene as the endogenous control to normalize the threshold cycle (CT).
Taqman universal pcr master mix and probes
Manufactured by Thermo Fisher Scientific
The TaqMan Universal PCR Master Mix and Probes are a reagent kit designed for use in real-time PCR experiments. The master mix contains the necessary components for DNA amplification and detection, while the probes provide target-specific fluorescent labeling. This product enables sensitive and specific quantification of target DNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy mini kit, by Qiagen (1 mentions)
Taqman reverse transcription reagent, by Thermo Fisher Scientific (1 mentions)
Gotaq qpcr master mix, by Promega (1 mentions)
Abi prism 7500 sequence detection system, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Anti rankl, by Bioss Antibodies (1 mentions)
Anti rank, by Bioss Antibodies (1 mentions)
Anti opg, by Bioss Antibodies (1 mentions)
Anti erk, by Bioss Antibodies (1 mentions)
Anti nf κb, by Bioss Antibodies (1 mentions)
Oligonucleotide primers, by Sangon (1 mentions)
Nanodrop, by Thermo Fisher Scientific (1 mentions)
Rneasy kit, by Qiagen (1 mentions)
3 protocols using taqman universal pcr master mix and probes
1
Gene Expression Analysis of Metabolic Pathways
2
VIP-Induced Osteoclastogenesis Regulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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VIP was purchased from Nanjing Peptide Biotechnology Inc. (Nanjing, China). Fetal bovine serum (FBS), a modification of Minimum Essential Medium (a-MEM), and L-glutamine were supplied by Invitrogen (Carlsbad, USA). Oligonucleotide primers were ordered from Sangon Biotech (Shanghai, China). TaqMan Universal PCR Master Mix and probes were obtained from Applied Biosystems (Foster City, CA). anti-RANKL, anti-RANK, anti-OPG, anti-NF-κB, anti-interleukin 6 (IL-6), anti-ERK, anti-carbonic anhydrase II (CAII), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) primary, and anti-IgG-horseradish peroxidase secondary antibodies were purchased from Bioss Biotechnology, Inc. (Beijing, China).
3
Aorta, Heart, Kidney, Adrenal RNA Profiling
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