LIANTI

The coverage in single cell methylation sequencing was poor, and the need arose to improve the coverage, accuracy and the read length of single cell. In our method, we found the most popular method only have 10% coverage in the 2x sequencing depth. Even if the most methods focused on the single strand library construction, and they ignored the base unblanced problem in the amplification. We found the DNA polymerase have amplification bias on the ATGC, so we used T7 transcription amplification to avoid the base unbalance in the amplifcaiton. The transposon insert the T7 adaptor in the genome without using random primer could increase the accuracy and tagmentation effiency. Then the gentle and high convertion efficiency kit is used by protect the methylation site by TET2 and convert the unmethylatate sites to the uracil by the APOBEC3A. Finally, we achieve the superior 70% coverage efficiency with only 2x sequencing depth on the single cell sequencing, which is the highest reported single cell methylation coverage until now.