Effects of different processing methods of human amniotic membrane on the quality of extracted RNA

ABSTRACT

The aim of this study was to determine the efficiency of different human amniotic membrane (HAM) processing methods on the concentration, purity and integrity of RNA. Two different techniques (Technique 1 and Technique 2) were employed for the processing of HAM, which differed in terms of washing solution, sample storage conditions and processing time. Based on preservation of HAM, three groups were formed under each technique. In Technique 1, the groups were fresh frozen 1 (F1), glycerol preserved (GP) and gamma irradiated glycerol preserved (IGP); where else in Technique 2, the groups were fresh frozen 2 (F2), 50% glycerol/Dulbecco’s modified Eagle medium (DMEM) cryopreserved HAM diluted with phosphate buffered saline (GB) and 50% glycerol/DMEM cryopreserved HAM diluted with diethylprocarbonate water (GD). Total RNA was extracted from the samples and their concentration, purity and integrity were examined. The F2 sample of which there was no pre-washing step and involved direct sample storage at -80ºC, shorter processing time and chilled processing conditions had yielded better quality of RNA compared to the others.