title: Gastric_Antrum_1
organism: Homo sapiens
description: Protocols: Fresh tissue sample for ultra deep sequencing was obtained from a gastroscopic biopsy (0.4 mm3). The patient was 33 years old, with no evidence of cancer and a normal gastroesophageal junction. Histological examination confirmed normal and healthy conditions. The total small RNA was collected from the tissue samples using the mirVana Isolation Kit (Ambion Inc., US). The concentration and quality were determined using a Nanodrop 1000 spectrophotometer (ND-1000; Nanodrop Technologies, Wilmington, DE), and purification and size selection were performed using 6% polyacrylamide gel electrophoresis Using the SOLiD Small RNA Expression Kit (Ambion Inc., US), 200 ng of small RNA (150-200 bp) was used as a template for the miRNA library. The miRNAs in the library were tagged with unique and specific amplification primers (The barcode system Life Technologies, CA, US). Next, 50 pg of the library was pooled with seven additional miRNA libraries at the same concentration. A fraction of the library pool (0.1 pg) was amplified and fixed on magnetic beads using emulsion PCR. The ePCR product was deposited onto a single slide, and multiplex SOLiD sequencing was used.