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Material Is No Longer Being a Matter for Methylation With A New Method

Andrew Adey and Jay Shendure from University of Washington updated a new method---Tn5mC-seq for detecting hydrogen sulfite salt. 1 ng of input DNA is needed for this method, which can still allow the whole-genome bisulfite sequencing. Complex bisulfite sequencing libraries generated by them are the core theory of this method.

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Material Is No Longer Being a Matter for Methylation With A New Method

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  1. Material Is No Longer Being a Matter for Methylation With A New Method • Andrew Adey and Jay Shendure from University of Washington updated a new method---Tn5mC-seq for detecting hydrogen sulfite salt. 1 ng of input DNA is needed for this method, which can still allow the whole-genome bisulfite sequencing. Complex bisulfite sequencing libraries generated by them are the core theory of this method.

  2. As for the investigation of genome-scale DNA methylation, there are alternatives, such as the whole-genome bisulfite sequencing. Although with the advantages of high-resolution, full methylation pattern detection, this method needs a majority of starting material. At least 5mg of genomic DNA is needed for the library construction. So this method is prohibitive for the samples with limited starting material. • Compared with this method, Low representative bisulfite sequencing needs less starting DNA, but comprehensive can not be achieved. This method is focusing on the specific area of the genome, which differs from the analysis of the whole genome. For researchers working on cancer and evolution, they have limited method selection with limited starting material. • Andrew Adey and Jay Shendure published the tagmentation-based whole-genome bisulfite sequencing on Genome Research. They utilized Tn5 transposase to DNA fragmentation and the incorporation of the joint. Compared with connection method, Tn5 transposase is more efficient and needs less starting DNA material.

  3. The previous cooperation between Jay Shendure and BGI facilitated a similar tagmentation-based library construction method for genome sequencing. They found this method can still offer high-quality coverage with less DNA. • Researchers updated previous method through attacking genomic DNA with transposon with single methylation firstly. And then with the oligonucleotide replacement strategy, they add the second methylation joint by annealing and fix the gap. And the 5th and 3rd ends of each chain have joints. Finally, they converted unmethylated cytosine to uracil through nitrite process. • In order to detect the preparation of transposon library, transposition and connections are applied to analyze a lymphoblastoid cell line with different content of the starting material. Finally, they found that although the content of starting material is greatly different, these two methods are relatively consistent. • This method offers a new selection for researchers working on epigenetics, such as the methylation research on cancer.

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