1. Briefly explain why urea is strongly preffered to NaOH for denaturation of tR
ID: 260662 • Letter: 1
Question
1. Briefly explain why urea is strongly preffered to NaOH for denaturation of tRNAs.
2. In an agarose gel containing ethidium bromide, rRNA can be located by its fluorescence, but mRNA cannot. Briefly explain.
3. Early experiments aimed at sequencing DNA were designed to mimic approaches used for proteins. Enzymatic end group analysis was a particularly popular method because wide variety of nucleases was available. However, while end group analysis with enzymes like aminopeptidase provided sequence information for the first 20 to 30 amino acids of a protein, end group analysis with nucleases like spleen phosphodiesterase failed to reach even those modest lengths. Briefly explain why nuclease-based end group anaylsis is so limited.
Explanation / Answer
1. Briefly explain why urea is strongly preffered to NaOH for denaturation of tRNAs.
Urea has strong interactions with the large greater surface areas that the bases have. Urea induced protein denaturation provides solubilisation of the bases, that leads to the driving force for denaturation. It changes the solubilisation of the proteins leading to denaturation. NaOH would be solubilising the proteins so only smaller peptides would be visible and as a result, it’s origin could not be traced. While using urea, there is only unfolding of the structures and not complete denaturation.
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