Rna Extraction Sample Clauses

Rna Extraction. Total RNA was extracted using Phase Lock Gel Heavy tubes (5 prime, VWR, Leicestershire, UK) by adding 200 μl of Chloroform per 1000 μl of Trizol reagent, shaking vigorously followed by 5-15 minutes incubation at room temperature. The solution was then centrifuged for 5-15 minutes at 12000 rpm at 4°C to separate the aqueous phase. To precipitate RNA 500 µl isopropanol per 1000 µl aqueous part was added to Eppendorf tubes, and again incubated for 5-10 minutes at room temperature. To obtain the total RNA pellet the solution was centrifuged for 10-15 minutes at 4°C and 12000 rpm. After discarding the aqueous part, 1ml of 75% ethanol was added to resuspend the pellet. The tubes were centrifuged for 5 minutes at 7300 g and then ethanol was poured away gently, and the pellet was air dried. Pellets were dissolved in 14 μl RNase free water (Qiagen). xxXXX quality and quantity analysed using Nano Drop 1000 Mass spectrometer (Xxxxxx Scientific). Ratios A260/A280 1.8–2 were considered to be of high purity and used for cDNA synthesis.

Rna Extraction. RNA was extracted from cells using Trizol® reagent, a solution of phenol and guanidine isothiocyanate.

Rna Extraction