I have recently been attempting to isolate RNA from polymer scaffolds using the Dynabeads mRNA DIRECT Purification Kit. When I measure using a nanodrop like I would for other extraction methods the RNA content is very low (1-2ng/uL). I assumed that this is because Dynabeads only extract the mRNA rather than total RNA as I still got amplification when carrying out cDNA conversion and qPCR, however I am wondering if there are any other methods for quantifying actual mRNA quantity?
You should see if anywhere at your institution has a Qubit fluorometer, as this will be the most accurate and sensitive way to quantify low RNA concentrations. You would need the Qubit high sensitivity RNA reagents from thermo fisher.
Yes, it is possible to measure RNA content when isolated using a DynaBeads mRNA direct kit. The DynaBeads mRNA direct kit is a kit for isolating and purifying mRNA from cells or tissue samples. After isolating the mRNA, you can measure its concentration using a spectrophotometer, which uses light absorption to measure the concentration of a substance in a solution. There are several methods for measuring RNA concentration using a spectrophotometer, including the absorbance at 260 nm (A260) and the absorbance at 280 nm (A280). The A260 method measures the amount of nucleic acid present in a sample, while the A280 method measures the amount of protein present in a sample. The A260/A280 ratio is often used to assess the purity of RNA samples.
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