Next, real time PCR was carried out in a final volume of 20 μl containing cDNA template, TS, DPD or GAPDH primers (Applied Biosystems, Tokyo, Japan) and TaqMan® Universal PCR Master Mix Applied Biosystemss, Tokyo, Japan), and DNase RNAase free water, using a StepOne real time PCR system (Applied Biosystems, Tokyo, Japan) according to the manufacturer's protocol.
One box filter is used to remove the noise outside the spectra band and reduce the data rate of the complex sampling for the next real-time computations.
To quantify CDK6 mRNA, 1 µg of total RNA was reverse-transcribed to cDNA using oligo dT and AMV reverse transcriptase (TaKaRa) in the reaction, which was performed with the following conditions: 42°C for 60 min and 70°C for 10 min. Next, real-time PCR was performed using the RT product, SYBER Green Dye (Invitrogen), and specific primers for CDK6 and GAPDH.
Next, real-time PCR was conducted on all DNA samples extracted from infected CEKs.
Next, real-time PCR was performed and methylation status was calculated by subtraction of Ct values.
Next, real-time RT-PCR was used to evaluate the mRNA levels of genes involved in the classic GSIS pathway.
Next, real-time PCR was performed using the RT product, SYBER Green Dye (Invitrogen) and specific primers for FEAT and GAPDH.
Next, real-time changes in the YFP/CFP emission ratio of the AKAR-18RBS reporter were monitored in cells expressing equivalent levels of the individual RIIα forms.
Next, real-time RT PCR analysis was performed using an Exicycler Quantitative Thermal Block (BiONEER, Daejeon, Korea) to quantitatively measure TCF4 mRNA levels from cell line samples or from clinical samples.
Next, real-time polymerase chain reaction (RT-PCR) was carried out in a final volume of 20 μL containing cDNA template, TS or GAPDH primers (Applied Biosystems, Tokyo, Japan), TaqMan® Universal PCR Master Mix Applied Biosystemss, Tokyo, Japan), and DNase RNAase free water, using the StepOne real-time PCR system (Applied Biosystems, Tokyo, Japan) according to the manufacturer's protocol.
Digital Image Processing Domain may be the old but the techniques and algorithms which are generating day by day are the trends of next generation real time applications.
