The protocol for an RNA concentration assay calls for a calibration curve with 15 ng/µL RNA in well #1, 150 ng/µL in well #2 and 550 ng/µL in well #3. An RNA standard has a concentration of 11.2 µg/µL. TE buffer is used to dilute the RNA to the appropriate concentrations.
A. If the standard was diluted by a factor of 10, what is its final concentration?
B. If a 200 µL solution of the standard was diluted with 300 µL of TE buffer what is the final concentration and what is the dilution factor (>1)?
C. How much of the RNA standard and TE buffer would you add in each well to give the appropriate final concentration with a final volume of 280 µL?
D. The assay kit contains a fluorescent dye that lists its concentration as "10x". How much dye should you add to get a final 1x concentration in each 300 µL well? Will you need to add buffer to reach a final volume of 300 µL? Or do you need to remove some assay from each of the three wells (if you need to, how much)?