I'm trying to determine amount of RNA extracted from zebrafish embryos (using Qiagen micro kit) and whether it is sufficient for cDNA/qPCR. Based on nanodrop output I appear to have an ongoing contamination issue and wondering about interpreting my RNA concentration and possible next steps. It seemed like the initial sample peak at the 225 range was removed after i precipitated the sample with ethanol/sodium acetate/glycogen, and on reanalyzing the sample the concentration of RNA has increased but 260/230 ratio is still low and 260/280 has remained at around 1.5. Is there a role here for a second precipitation or other options, or would simply trying a new sample be advisable.
Sorry, but neither spectrum looks like a decent prep of nucleic acid, which should display a prominent peak at 260 nm. Thus, it is impossible to gauge the RNA concentration in your sample.The strong absorbance at 230 nm is likely due to contamination by thiocyanate, which is present in high concentration in the lysis buffer. While performing spin column purification, it is advisable to perform more than one EtOH wash and to wipe the outside of the spin column with a Kleenex between washes. I would start the prep over again, if getting the source material is not too difficult; alternatively, you could try passing the prep again over a spin column after adding buffer to it and going again through the whole purification process.
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