Table 1: Reaction setup.

Setup and conditions for poly-C, PCR 1and PCR 2reactions.

Reaction	Setup	Conditions
Poly-C reaction	30 μL of sheared gDNA	Incubate at 37°C for 1 hour
	2.5 μL of 9.5 mM dCTP/0.5 mM ddCTP
	10 μL of 5X Terminal deoxynucleotidyl transferase (Tdt) reaction buffer
	1.25 μL of rTdt
	6.25 μL of water to 50 μL
PCR 1	23 μL 3’-poly-C purified DNA (entire sample from previous step)	1 cycle: 2 min 94 °C
		15 cycles: 15 s 94 °C
	10 μL 10x high-fidelity DNA	30 s 60 °C
	polymerase reaction mix	2 min 68 °C
	2 μL 10mM dNTPs	1 cycle: 4 min 68 °C
	2 μL 50 mM MgS04	Hold: ∞ 4 °C
	1 μL 30 μL 510 biotin
	3 μL 30 μL olj 376
	0.5 μL high-fidelity DNA polymerase
	8.5 μL pure water to 50 μL total
PCR 2	DNA bound beads from previous step	1 cycle: 2 min 94 °C
	10 μL 10x high-fidelity DNA	15 cycles: 15 s 94 °C
	polymerase reaction mix	30 s 60 °C
	2 μL 10mM dNTP	2 min 68 °C
	2 μL 50 mM MgSO4	1 cycle: 4 min 68 °C
	1 μL 30 μM olj 511	Hold: ∞ 4 °C
	1 μL 30 μM barcode primer (Table 2)
	0.5 μL high-fidelity DNA polymerase
	33.5 μL pure water to 50 μL