Journal: International Journal of Chronic Obstructive Pulmonary Disease

Article Title: Peripheral leukocyte microRNAs as novel biomarkers for COPD

doi: 10.2147/COPD.S130416

Figure Lengend Snippet: The description of small RNA sequencing data

Article Snippet: Adaptor ligation: We prepared a connection 3′adaptor system (TruSeq Small RNA Sample Pre Kit, Illumina, San Diego, CA, USA) utilizing the following reaction conditions: 70°C for 2 min; 28°C for 1 h; 28°C for 15 min. Then, we added the 5′adaptor mix system with the following reaction conditions: 70°C for 2 min; 28°C for 1 h. RT-PCR: We prepared the first strand master mix and super script II (Invitrogen) reverse transcription (reaction condition: 70°C for 2 min; 50°C for 1 h); this was followed by several rounds of PCR amplification with PCR primer cocktail and PCR master mix to enrich the cDNA fragments (reaction conditions: 98°C for 30 s; 11 cycles of 98°C for 10 s, 60°C for 30 s, 72°C for 15 s; 72°C for 10 min; 4°C hold).

Techniques: RNA Sequencing, Control