Rapid, high-quality and epidermal-specific isolation of RNA from human skin

Abstract

As global transcriptome analyses with a growing demand on layer-specific applications are widely used in cutaneous biology, we investigated the effect of established and optimized dermo-epidermal separation methods on the quality of RNA. We compared enzymatic separation with dispase, chemical separation with 1 m sodium chloride and heat separation to a treatment with 3.8% ammonium thioyanate. The impact of freezing as well as the addition of 10m m aurintricarboxylic acid was considered in the evaluation of the amount and quality of isolated RNA from dermis and epidermis. Using the low abundant gene kallikrein 12 for real-time PCR analysis, we were able to demonstrate the superior RNA quality after dermo-epidermal separation using 3.8% ammonium thiocyanate. In addition to the time effectiveness this separation technique promises dermal and epidermal purity and is therefore the method of choice for producing high-quality RNA for genome-wide dermal and epidermal transcriptional analysis. © 2007 The Authors Journal compilation © 2007 Blackwell Munksgaard.