Oligo dt random primers and reverse transcriptase

Total RNA was extracted from the small intestines using TRIzol Reagent (Invitrogen). A 10 μg aliquot of each RNA sample was reverse transcribed into cDNA using oligo-dT random primers and reverse transcriptase (Takara, Dalian, China), and RT-PCR was performed using SYBR Premix Ex TaqTM II (Takara). Primers used were as follows:
Wnt3a: 5′-TTCTTACTTGAGGGCGGAGA-3′ (forward) and 5′-ACCCGTATCCCAGACAGGA-3′ (reverse); IGF-1: 5′-CAACTCCCAGCTGTGCAATT-3′ (forward) and 5′-GCCGAGGTGAACACAAAACT-3′ (reverse); HGF: 5′-CAGGACCATGTGAGGGAGAT-3′ (forward) and 5′-TACCAGGACGATTTGGGATG-3′ (reverse); bFGF: 5′-AAGGGAGTGTGTGCCAACC-3′ (forward) and 5′-GCCCAGTTCGTTTCAGTGC-3′ (reverse); VEGF: 5′-CCCTTCGTCCTCTCCTTACC-3′ (forward) and 5′-AAGCCACTCACACACACAGC-3′ (reverse); TGF-β1: 5′-ATTCCTGGCGTTACCTTGG-3′ (forward) and 5′-AGCCCTGTATTCCGTCTCCT-3′ (reverse); Lgr5: 5′-CGGAGGAAGCGCTACAGAAT-3′ (forward) and 5′-CTGGGTGGCACGTAGCTGAT-3′ (reverse); and GAPDH: 5′-GGTGGGTGGTCCAAGGTTTC-3′ (forward) and 5′-TGGTTTGACAATGAATACGGCTAC-3′ (reverse).

Total RNA was extracted from the small intestines using TRIzol Reagent according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA). A reverse transcription reaction was performed with 10 μg aliquot of each RNA sample using oligo-dT random primers and reverse transcriptase (Takara, Dalian, China), and real-time PCR was performed using a SYBR Premix Ex TaqTM II (Takara). Primer sequences were as follows:
Bcl-2: 5′-AGGATTGTGGCCTTCTTTGA-3′ (forward) and 5′-CAGATGCCGGTTCAGGTACT-3′ (reverse); XIAP: 5′-TGGAGACTCAGCTGTTGGAA-3′ (forward) and 5′-CTGGCCATTTTGGATACCAG-3′ (reverse); uPA: 5′-CGCACACTGCTTCATTCAAC-3′ (forward) and 5′-GCTGCTCCACCTCAAACTTC-3′ (reverse).