Cells were collected into trizol® reagent (thermofisher) and total RNA was extracted using ‘Pureling RNA mini kit’ (thermofisher) according to manufacturer’s instructions. 5ug of total RNA was used in reverse transcriptase polymerase chain reaction performed using ‘RNA to cDNA Ecodry™ premix (oligo dT)’ cDNA synthesis kit (Clontech) according to manufacturers instructions. Resulting cDNA corresponding to 50ng of total RNA was used in each 20ul of quantitative real time PCR reaction. qRT-PCR was performed using SybrGreen master mix (Biorad) and the relative expression of each gene was calculated after normalizing to β-Actin endogenous control and using comparative ΔCt method. A list of the primers used is in Supplementary Table 4 .
Rna to cdna ecodry premix oligo dt cdna synthesis kit
Manufactured by Takara Bio
The RNA to cDNA Ecodry™ premix (oligo dT) cDNA synthesis kit is a reagent used for the conversion of RNA to complementary DNA (cDNA). It contains a premixed solution with the necessary components for the reverse transcription reaction, including a reverse transcriptase enzyme and oligo(dT) primers.
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2 protocols using rna to cdna ecodry premix oligo dt cdna synthesis kit
1
RNA Extraction and qRT-PCR Analysis
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RNA Extraction and qRT-PCR Analysis
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Cells were collected into trizol® reagent (thermofisher) and total RNA was extracted using ‘Pureling RNA mini kit’ (thermofisher) according to manufacturer’s instructions. 5ug of total RNA was used in reverse transcriptase polymerase chain reaction performed using ‘RNA to cDNA Ecodry™ premix (oligo dT)’ cDNA synthesis kit (Clontech) according to manufacturers instructions. Resulting cDNA corresponding to 50ng of total RNA was used in each 20ul of quantitative real time PCR reaction. qRT-PCR was performed using SybrGreen master mix (Biorad) and the relative expression of each gene was calculated after normalizing to β-Actin endogenous control and using comparative ΔCt method. A list of the primers used is in Supplementary Table 4 .
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