Human miFinder miRNA PCR array of liver biospy specimen of chronic hepatitis C virus infected patients

The miRNAs were purified from paraffin embedded liver biopsy samples of HCV and HCC patients, using an miRNeasy mini kit (Qiagen) according to manufacturer’s instructions. First, 25mg of liver biopsy tissues was homogenized in 700μl QIAzol lysis reagent, mixed, and shaken with chloroform. After that, the phases were separated by centrifugation at 12000´g for 15 min. The upper aqueous phase was collected in a fresh tube and the remaining DNA and organic protein organic phases was stored at -70°C until required. The RNA aqueous phase was washed further on the column and finally RNA was eluted into DEPC water (Ambion). Purified miRNAs were reverse transcribed into first strand cDNA using a miRNA first strand kit. (Qiagen). The reaction was carried out at 37ºC for 2 hours followed by heating at 95ºC for 5 min. The liver biopsies classified as fibrosis stage F1 and F2 were grouped together and named as early HCV (E-HCV, n=4), while fibrosis stage F3 and F4 were grouped together and named as advanced fibrosis (A-HCV, n=5).