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本帖最后由 细胞海洋 于 2012-2-8 01:20 编辑
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0 V3 B8 O7 F. m1 s! f! g/ W3 [ 不得不承认，现在我们被美国人牵着鼻子走，上个世纪60年代的时候，美国人提出了甲基化，国内还不知道基因组，现在我们正在用大把大把的银子做全基因组甲基化的时候，RNA的甲基化检测又被人家想出来了，总之想法总是人家的。4 g1 h1 k$ M _5 E
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1， Detection of cytosine methylation in RNA using bisulfite sequencing : i6 G; O1 b) `0 k
/ K. a; v: L% I; u. R. t: e2， RNA methylation and control of eukaryotic RNA biosynthesis.
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Post-transcriptional RNA modifications are a characteristic feature of noncoding RNAs and have been described for ribosomal RNAs (rRNAs), transfer RNAs (tRNAs), and various other small RNAs. However, the biological function of most of these modifications remains uncharacterized. Cytosine-5 methylation (5mC) has been detected in abundant and long-lived RNA molecules such as rRNAs and tRNAs, but, because of technical limitations, the occurrence of base-methylated cytosines in other RNAs is not known. To facilitate the detection of RNA methylation, we have established a method for analyzing base-methylated cytosines in RNA using bisulfite sequencing. Treatment of RNA with bisulfite causes the chemical deamination of nonmethylated cytosines to uracil, while methylated cytosines remain unaffected. cDNA synthesis followed by polymerase chain reaction (PCR) amplification and DNA sequencing allows investigators to reproducibly and quantitatively distinguish unmethylated cytosines (as thymines) from methylated cytosines in tRNAs and rRNAs. Using high-throughput sequencing approaches, this protocol should enable the characterization of 5mC methylation patterns in any RNA molecule, including low abundance RNAs. ; P. h; V% k& E Q# i8 }( i) m: p
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4楼原文 感谢susanuse 提供 |
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发表于 2012-2-7 15:25
发表于 2012-2-7 15:32
发表于 2012-2-8 01:19
