Real-time quantitative PCR analysis of murine non-ischemic and ischemic wound edge

Wound edges from a murine bipedicle flap ischemic wound model were collected 3 days post wounding. RNA from ischemic and non-ischemic wound edges was isolated by miRVana RNA isolation kit. Mature miRNAs were then profiled by real-time PCR. Wound edges were collected from ischemic (bipedicle flap model) and non-ischemic wounds made in C57Bl6 mice on 3 days post-wounding. RNA was isolated by miRVana RNA isolation kit. Mature miRNAs were profiled by real-time PCR using an Applied Biosystems 7900HT real-time PCR instrument equipped with a 384 well reaction plate. RNA (500 ng) was converted to cDNA by priming with a mixture of looped primers (Mega Plex kit, Applied Biosystems, Foster City, CA). Liquid-handling robots and the Zymak Twister robot were used to increase throughput and reduce error. Real-time PCR was performed using standard conditions