1/4/2024 0 Comments Dot blot![]() Li et al., 2017), methyl-sensitivity of MazF RNA endonucleases ( Semi-quantitative methods include dot blot technology ( Z. Currently, a variety of methods have been developed to identify mĪ modifications in cells, which can be divided into three categories: semi-quantitative, quantitative, and precise location detection. Also, detection of mĪ is important for studying its biological functions and mechanisms. The correlation between the level of mĪ modification and clinicopathological features has been shown in diverse tumors, which may provide prognostic value in these diseases.Ī modification in vitro can help identify the precise regulatory forms and synergistic roles of mĪ modifications in cancer and other diseases. In mammals, mĪ methylation plays a variety of key roles including embryonic development, neurogenesis, circadian rhythm, stress responses, sex determination, and tumorigenesis (Ī modification is vital during stem cell proliferation, with METT元 depletion reducing the differentiation of embryonic stem cells ( ), and DNA damage ( Xiang et al., 2017 E. With methyltransferase activity consist of three individual proteins: METT元, METTL14, and WTAP (Ī modification to modulate the fate of mRNA (Ī plays a pivotal role during various biological processes including virus infection ( This protocol can also be used to enrich specific RNAs (such as tRNA, rRNA, or miRNA) by isolation technology to detect the mĪ level of single RNA species, so as to facilitate further studies of the role of mĪ) is the most prevalent internal RNA modification in eukaryotic mRNAs and long non-coding RNAs (Ī modification refers to the methylation of the nitrogen atom at position 6 of the RNA adenosine, mainly located on the common motif of RRmĪCH (R denotes A or G, H denotes A, C, or U) (Ī modification is regulated by the combined action of methylases and demethylases ( Here, we describe in detail the dot blotting method for detecting mĪ levels in RNA (mRNA as an example), including total RNA extraction, mRNA purification, dot blotting, and data analysis. ![]() Common mĪ methylation detection techniques play an important role in understanding the biological function and potential mechanism of mĪ, mainly including the quantification and specific localization of mĪ modification sites. The mĪ modification is also extensively present in non-coding RNAs, including microRNAs (miRNAs), ribosomal RNAs (rRNAs), and transfer RNAs (tRNAs). ![]() A) is the most prevalent internal modification of eukaryotic messenger RNAs (mRNAs), affecting their fold, stability, degradation, and cellular interaction(s) and implicating them in processes such as splicing, translation, export, and decay. ![]()
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