Express photolithographic DNA microarray synthesis with optimized chemistry and high-efficiency photolabile groups

Table 2 Representative chemical synthesis protocol, in Expedite 8909 format, used for the legacy syntheses

Cycle NPPOC-dT (legacy synthesis)
Function	Mode	Pulses	Sec	Description
$Coupling
1/*Wsh	Pulse	20	0	Flush with Wsh
2/*Act	Pulse	6	0	Act
21/*T + Act	Pulse	5	0	T + Act
2/*Act	Pulse	6	0	Push with Act
1/*Wsh	Pulse	3	60	Couple monomer
1/*Wsh	Pulse	10	0	Flush with Wsh
$Capping
40/*Gas A	Pulse	1	30	Dry column
$Oxidizing
15/*Ox	Pulse	30	0	Ox to column
12/*Wsh A	Pulse	45	0	Flush with Wsh A
17/*Aux	Pulse	15	0	Exposure solvent
130/*Event 2 Out	Na	4	1	Event 2 out
17/*Aux	Pulse	10	50	Exposure solvent
12/*Wsh A	Pulse	5	20	Flush with Wsh A
130/*Event 2	Na	4	1	Event 2 out
12/*Wsh A	Pulse	20	0	Flush with Wsh A

The 5′-NPPOC light-deprotection step takes place between the two “Event Out” commands, which trigger the opening and closing of a shutter blocking the UV light. The 70 s exposure, at 80 mW/cm², corresponds to an exposure of 5.6 J/cm². The oxidation, the first two steps after the $Oxidizing header, are used intermittently (every ~40 cycles) and after the last coupling

ISSN: 1477-3155