Fig. 1.

Schematic overview of SRSLY. A DNA input pool of diverse template molecules is denatured with heat and maintained as single-stranded molecules through a cold-snap and use of a thermostable single-stranded DNA binding protein (SSB). Template DNA is phosphorylated and SRSLY splint adapters are ligated in a combined phosphorylation/ligation reaction. Adapters contain a random single-stranded splint overhang and ligation blocking modifications on all termini except for the ones that facilitate correctly oriented library molecules. After clean up, molecules are ready for index PCR