Nanodrop nd 1000 uv vis spectophotometer

Frozen cervical tissue immersed in TRIzol reagent was homogenized using a homogenizer (Bio-gen Pro200 Homogenizer, Pro Scientific) in order to lyse the tissue. The RNA extraction was completed using the RNeasy Kit (Quiagen Ltd., Crawley, West Sussex, UK) according to the manufacturer’s instructions. Total RNA concentration was quantified using the Nanodrop ND-1000 UV-Vis Spectophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA). Quality of RNA was ascertained with the use of 2100 Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). RNA integrity number (RIN) was greater than 7 in all samples and RNA aliquots were stored at − 80 °C after extraction.

In order to lyse the tissue and extract the RNA, frozen cervical tissue was immersed in TRIzol reagent and then homogenized using the homogenizer (Bio-gen Pro200 Homogenizer, Pro Scientific). The RNA extraction was completed using the RNeasy Kit (Quiagen Ltd., Crawley, West Sussex, UK) according to the manufacturer’s instructions. Total RNA concentration was quantified using the Nanodrop ND-1000 UV-Vis Spectophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA). Quality of RNA was ascertained with the use of 2100 Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). RNA integrity number (RIN) was greater than 7 in all samples and RNA aliquots were frozen at − 80 °C after extraction.

Endometrial cytobrushes were placed in 350 µL of TRIzol reagent and vortexed vigorously for 1 min to lyse the cells. Following incubation at room temperature for 5 min, chloroform was added to each sample, shaken vigorously for 15 s and centrifuged at 12,000×g for 15 min at 4 °C. The aqueous phase was transferred to a new tube. Ethanol (70%) was added to the aqueous phase, mixed immediately, and then transferred to an RNeasy spin column. The RNA extraction was completed using the RNeasy kit (Qiagen Ltd., Crawley, UK), following the manufacturer’s instructions. This included a number of wash steps using the kit buffers supplied, and finally elution in 30 µL of RNase-free water. The quantity and quality of RNA present in the samples was determined using the NanoDrop ND-1000 UV–Vis Spectophotometer (NanoDrop Technologies Inc., Wilmington, DE, USA) and 2100 Agilent Bioanalyzer respectively.