Go taq flexi dna polymerase

Cells treated with arecoline for specific periods of time and biopsy samples from patients (n = 5) were used for RT-PCR analysis. Total RNA was purified from samples using TRIzol^® according to the manufacturer’s instructions. For mRNA analysis, complementary DNA of each sample was randomly primed from 1 μg of total RNA using Superscript-RT. Real time PCR was performed using the KAPA SYBRFAST qPCR KIT. Data was normalized to18s rRNA and relative expression levels were determined using StepOne Real Time PCR software. Semi-quantitative PCR analysis was carried out in a total volume of 10 μl containing 0.5 picomoles of each primer using Go Taq Flexi DNA Polymerase on the 2720 Thermal Cycler (Applied Biosystems, USA) The relative quantification value for each target gene was expressed as 2^−ΔΔCT [49 (link)].

For the PCR reaction, a ProFlex PCR System thermocycler (Applied Biosystems, MA, USA) and GoTaq® Flexi DNA polymerase were used (Madison, USA), following the manufacturer's guidelines. The amplification products were revealed by horizontal electrophoresis using a subcell electrophoresis chamber (Bio-Rad, CA, USA) on a 2% agarose gel stained with HydraGreen™ (ACTGene, NJ, USA) and a molecular weight marker of 100 bp (New England Biolabs, USA). The gel was visualized and documented using the ENDURO GDS Gel Documentation System (Labnet International, NJ, USA). The amplification products were purified and sequenced by the Sanger method (Macrogen Inc., Korea).