Seq-Well S3
Seq-Well S3 modified the following protocol steps from v1, above (Hughes et al., 2019). First, hybridization buffer was supplanted with 8% (v/v) polyethylene glycol (PEG, Sigma). Second, after exonuclease digestion, bead-associated cDNA was denatured for 5 min in 0.2 mM NaOH with end over end rotation. Next, beads were washed with TE + 0.01% tween-20, and second strand synthesis was carried out by resuspending beads in a master mix containing Klenow Fragment (NEB), dNTPs, PEG, and the dN-SMRT oligonucleotide to enable random priming off of the beads.