Total RNA (500 ng) was used for reverse transcription. Reverse transcription was carried out using ReverTra Ace (TOYOBO) following the manufacturer’s instructions. Reverse-transcribed cDNAs were subjected to realtime PCR, which was performed with a CFX96 Touch Real-Time PCR System (Bio-Rad). For the detection of CD44, Cdkn1a (Mm.PT.58.5884610), Mdm2 (Mm.PT.58.42166864), CXCL2 (Mm.PT.58.10456839), CD44 (Mm.PT.56a.33229033), a Prime Time Mini qPCR Assay from Integrated DNA Technologies was used. Custom-designed TaqMan Dual-Labeled Probes from Sigma were also used for the detection of p53, PHLDA3, IER5, RPRM, PMAIP, PTP4A1, FUCA1, PLK2 and β-actin. SYBR Premix Ex Taq II (Takara) probe was used for detection of Adam8 and mPGES1.
Taqman dual labeled probes
Manufactured by Merck Group
TaqMan Dual-Labeled Probes are oligonucleotide probes used in real-time PCR assays. They contain a reporter dye at the 5' end and a quencher dye at the 3' end. During the PCR process, the probe binds to the target sequence and is cleaved by the 5' exonuclease activity of the DNA polymerase, separating the reporter and quencher dyes, resulting in an increase in fluorescent signal.
Automatically generated - may contain errors
Lab products found in correlation
Revertra ace, by Toyobo (2 mentions)
Cfx96 touch real time pcr system, by Bio-Rad (2 mentions)
Prime time mini qpcr assay, by Integrated DNA Technologies (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
Superscript first strand synthesis system for rt pcr, by Thermo Fisher Scientific (1 mentions)
Taqman probe, by Integrated DNA Technologies (1 mentions)
Taqman probe, by Thermo Fisher Scientific (1 mentions)
Lightcycler 480 instrument, by Roche (1 mentions)
2 protocols using taqman dual labeled probes
1
Gene Expression Analysis by qPCR
2
Real-Time PCR Analysis of Heat Shock Genes
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Reverse transcription was carried out using the SuperScript First-Strand Synthesis System for RT-PCR (Life Technologies) or ReverTra Ace (TOYOBO) following the manufacturer’s instructions. Total RNA (0.2–5 μg) was used for reverse transcription. Reverse-transcribed cDNAs were subjected to real-time PCR, which was performed with a LightCycler 480 Instrument (Roche Diagnostics) or CFX96 Touch Real-Time PCR System (Bio-Rad). For the detection of IER5, HSF1, HSPA1A and GAPDH, a TaqMan probe (IER5; Hs00275419, HSF1; Hs00232134_m1, HSPA1A; Hs00359163_s1, GAPDH; Hs02758991_g1)from Applied Biosystems were used. For the detection of HSPA6 and HSPA1B, a TaqMan probe from Integrated DNA Technologies was used. In some cases, custom-designed TaqMan Dual-Labeled Probes from Sigma-Aldrich were also used for the detection of HSPA1A and GAPDH.
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