NextSeq 500 sequencing; A time-course study of thioacetamide (TAA)-induced liver fibrosis in rats
A time-course study of thioacetamide (TAA)-induced liver fibrosis in rats
To induce hepatic fibrosis, Male Sprague Dawley rats (9-12 weeks of age and 380-420 g of weight upon arrival, supplied by Beijing Vital River laboratory animal Co., Ltd.) were treated with thioacetamide (TAA). Rat liver samples were collected from five groups of rats at week 1, 2, 4, 8 and 13 after TAA (300 mg/kg) administration three times per week while five control groups receive the same volume of 0.9% normal saline. Four biological replicates were used for each group. Frozen rat liver tissue (~ 2 mm cube) was lysed and homogenized using Trizol reagent in a Qiagen TissueLyser II. After phase separation of the homogenate, the upper aqueous phase was subject to total RNA purification using Qiagen RNeasy Mini spin column with extensive on-column DNase I digestion. Purified total RNA samples were analyzed by Agilent Bioanalyzer RNA 6000 Nano assay and quantified by Qubit RNA assay.  For construction of sequencing libraries, 1 ug of total RNA (RIN > 8) was subject to Illumina TruSeq Stranded Total RNA Sample Prep kit with Ribo-Zero Gold. Libraries were analyzed by Bioanalyzer High Sensitivity DNA assay and quantified by Qubit DNA assay.
Experimental Factor: compound
none
Experimental Factor: disease
normal
Experimental Factor: time
8