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ssDRIP sequencing

QL Qin Li
JZ Jincong Zhou
SL Shuai Li
WZ Weifeng Zhang
YD Yingxue Du
KL Kuan Li
YW Yingxiang Wang
QS Qianwen Sun
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Seedlings grown on 1/2 MS for 5 days were transferred to MS liquid medium containing 1 μM DMSO or CPT for 2-h treatment, and genomic DNA was extracted from roots tips using Honda buffer without filtering as described in34,35. Then other steps followed the ssDRIP-seq protocol according to references34,35. In brief, 1 μg fragmented DNA (digested by DdeI (R0175S, NEB), MseI (R0525S, NEB), NlaIII (R0125S, NEB) and MboI (R0147S, NEB) endonucleases) was used for S9.6 immunoprecipitation. Meanwhile, the same amount of RNase H-treated fragmented DNA was used for S9.6 immunoprecipitation as the negative control. For each immunoprecipitation assay, 8 μg S9.6 antibody and 40 μL Protein G beads were used. The precipitated DNA was used for sequencing library constructions using an ssDNA library preparation kit (ND620, Vazyme).

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