Tn5 transposon tagments ssDNA. (A) Tn5 transposon tagmented genomic DNA (gDNA). gDNA from HEK293T cells was subjected to Tn5 transposon tagmentation. (B) Tn5 transposon had little activity to tagment total RNA. Total RNA purified from HEK293T cells was subjected to Tn5 tagmentation reaction. RNase A, which digested RNA, was used as the control for the existence of RNA. (C) Tn5 transposon tagmented both dsDNA and ssDNA. dsDNA and ssDNA, which were generated from asymmetric PCR, were used for Tn5 tagmentation. ExoI, which only digested ssDNA, was used to indicate the ssDNA. (D) dsDNA and ssDNA, which were generated from asymmetric PCR, were purified and treated with different nucleases. ExoI, which only digested ssDNA, was used to indicate the ssDNA. DNase I, which digested both ssDNA and dsDNA, was used to indicate the DNA. (E) Tn5 transposon could not tagment ssDNA <130 nt. Different lengths of ssDNA, which were generated from asymmetric PCR, were used for Tn5 tagmentation. (F) Tn5 transposon tagmented dsDNA and ssDNA with similar efficiencies. Purified dsDNA and ssDNA were subjected to Tn5 transposon tagmentation under different temperature for 5 min.
