MALAT1, PTENP1, EMT biomarkers genes (E-CADHERIN, N-CADHERIN, SLUG, SNAIL, ZEB1, TWIST, MMP-9, and VIMENTIN) and mRNA expression of 88 genes which involved in signaling pathways of colorectal tumorigenesis (ref KEGG) were analyzed in generated 5FU resistance cell lineages of, as well as pcDNA3.0-PTENP1 and si-MALAT1 transfected HT-29 cells using RT-PCR analysis. Total RNA was extracted from the cells with E.Z.N.A." Total RNA Kit I (Omega Bio-tek, Inc., Norcross, GA, USA). All RNA samples were assessed for quality using a spectrophotometer (Beckman Coulter, Inc., Fullerton, CA). ProtoScript First Strand cDNA Synthesis Kit (Biolabs, USA) in a thermal cycler (i-Cycler, Bio-Rad Laboratories, USA). Taqman probe-based RT-qPCR reaction was performed according to the manufacturer's protocol (Thermo Fisher, US) using Step One Plus Real-time PCR system (Applied Biosystems, USA). The analysis was carried out in three replicates.
Taqman probe based rt qpcr reaction
Manufactured by Thermo Fisher Scientific
Sourced in United States
The TaqMan probe-based RT-qPCR reaction is a real-time reverse transcription-quantitative polymerase chain reaction (RT-qPCR) technique. It utilizes a fluorescent probe to detect and quantify specific RNA targets during the amplification process.
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2 protocols using taqman probe based rt qpcr reaction
1
Molecular Profiling of 5FU Resistance
2
Molecular Profiling of 5FU Resistance
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MALAT1, PTENP1, EMT biomarkers genes (E-CADHERIN, N-CADHERIN, SLUG, SNAIL, ZEB1, TWIST, MMP-9, and VIMENTIN) and mRNA expression of 88 genes which involved in signaling pathways of colorectal tumorigenesis (ref KEGG) were analyzed in generated 5FU resistance cell lineages of, as well as pcDNA3.0-PTENP1 and si-MALAT1 transfected HT-29 cells using RT-PCR analysis. Total RNA was extracted from the cells with E.Z.N.A." Total RNA Kit I (Omega Bio-tek, Inc., Norcross, GA, USA). All RNA samples were assessed for quality using a spectrophotometer (Beckman Coulter, Inc., Fullerton, CA). ProtoScript First Strand cDNA Synthesis Kit (Biolabs, USA) in a thermal cycler (i-Cycler, Bio-Rad Laboratories, USA). Taqman probe-based RT-qPCR reaction was performed according to the manufacturer's protocol (Thermo Fisher, US) using Step One Plus Real-time PCR system (Applied Biosystems, USA). The analysis was carried out in three replicates.
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