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2 years ago
2 years ago by Paul G. Macon
Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?
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|Total RNA was extracted from the tissues according to the manufacturer’s instructions (Spin-column, BioTeke, Beijing, China). First-strand cDNA was obtained using PrimeScript First-strand cDNA synthesis kit with Oligo dT Primer from Takara (Dalian, China). The cDNA mixture was diluted 10 times with PCR-grade water and then stored at −80 °C for qRT-PCR analysis.|