Problem:
Question 1: Your RNA preparation is denatured by adding the FFMB mix (containing formamide and formaldehyde) and by heating to 90oC before loading the RNA into the gel. Why is it necessary to heat the samples to 90oC? Why are the denaturants in the FFMB mix needed?
Question 2: Does guanidinium isothiocyanate in the denaturing agarose gel denature the RNA molecules? Explain.
Question 3: The polar and non-polar phases from the phenol-chloroform extracted lysed cell suspension are separated in the PLG tubes. What property of the PLG gel is required to bring about this separation?
Could someone help to explain the answer of these questions.