What is Epigenetic?
Biological mechanisms in which DNA, RNA, and proteins are chemically or structurally modified, without changing their primary sequence (a.k.a mutation).
Epigenetic regulatory mechanisms include DNA methylation and
hydroxymethylation, histone modification, chromatin remodeling, RNA
methylation, and regulation by small and long non-coding RNAs.
What is RNA methylation?
Although the majority of nucleic acid modification research involves 5-methylcytosine (5-mC) in DNA, RNA is also extensively modified. In fact, there exist over a hundred modifications to RNA, including 5-mC, and RNA methylation is a common and naturally-occurring event in both prokaryotic and eukaryotic organisms. RNA methylation has been observed in numerous types of RNA molecules, including mRNAs, tRNAs, and non-coding RNAs, but the function of RNA methylation remains unclear.Source: Zymo
How to use EZ RNA Methylation Kit ?
Follow the following flowchart and the complete protocol here
Source: Zymo
What happened to RNA methylation?
Sequencing results following bisulfite treatment. RNA with methylated C (5-mC) at nucleotide position #7 was processed using the EZ RNA Methylation™ Kit.
The recovered RNA was amplified by RT-PCR and then cloned and
sequenced. The methylated cytosine at position #7 remained intact while
the non-methylated cytosines at positions #3, 4, and 13 were completely
converted into uracil (post-bisulfite treatment) and detected as thymine
following RT-PCR and sequencing
Source: Zymo
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