Fast start sybr green master kit

Total RNA was extracted from the cells using Trizol^® (Invitrogen, Carlsbad, CA, USA). One microgram of RNA was used for reverse transcriptase reactions. Gene expression was carried out using the Fast Start SYBR Green Master kit (Applied Biosystems, Foster City, CA, USA), using a 7500 Real-Time Thermal Cycler (Applied Biosystems). Relative gene expression values were normalized to the constitutive expression of the RPLP0 gene encoding 60S acidic ribosomal protein P0. Values were determined using the 2^−ΔΔCT method [26 (link)]. The specific primers for RPLP0 [4 (link)] and TP63 [27 (link)] are shown in Table S1 (Supplementary Materials).

Total RNA was extracted from the from colon tissue using the DNA/RNA/PROTEIN Kit purification plus (NORGEN BioTek Corp., Thorold, ON, Canada) following the manufacturer’s instructions. The RNA concentration was determined by measuring the absorbance at 260 nm. One microgram of RNA was used for first-strand cDNA synthesis with Revert Aid H Minus First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, Waltham, MA, USA). Gene expression was carried out using the Fast Start SYBR Green Master kit (Applied Biosystems, Foster City, CA, USA), using a 7500 Real-Time Thermal Cycler (Applied Biosystems). Relative gene expression values were normalized to the constitutive expression of the RPLP0 gene. Specific primers for target genes are shown in Supplementary Table S1.