Human pancreas total rna

Total RNA was isolated from cultured cells using NucleoSpin miRNA (TaKaRa, Otsu, Japan). RNA concentration and purity were assessed by UV spectrophotometry. RNA integrity was checked by formaldehyde gel electrophoresis. Total RNA from normal epithelial tissues from colon and pancreas were human colon total RNA and human pancreas total RNA (TaKaRa). To assess the expression levels of MIR143-3p, we conducted qRT-PCR using TaqMan miRNA assays (Applied Biosystems) and the Thunderbird Probe qPCR Mix (Toyobo, Osaka, Japan) according to the manufacturer’s protocol. RNU6B was used as an internal control. Relative expression levels were calculated using the ΔΔCt method.

Total RNA was isolated from differentiated cells using an RNeasy Micro Kit (QIAGEN, Venlo, the Netherlands), and human pancreas total RNA (Takara Bio USA, Mountain View, CA, USA) was used for the comparison of the expression of pancreatic genes. cDNA was synthesised using a High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Stockholm, Sweden) with random hexamer primers. Real-time PCR analysis of each sample was then performed with a StepOne and StepOnePlus Real-Time PCR System (Applied Biosystems). TaqMan assays for real-time PCR (Applied Biosystems) are listed in Supplementary Table 1.