Table 1.

SLiCE reaction conditions

Components	Standard cloning	Rush cloning	BAC cloning
Vector DNA	Purified, 50–200 ng	Unpurified, 1 μLa	Purified, 50–200 ng
Insert DNA	Purified, 1:1–10:1b	Unpurified, 1 μLa	Phenol/chloroform-purified, 5–10 μg restriction enzyme digested BAC fragments
10× SLiCE buffer	1 μL	1 μL	1 μL
PPY SLiCE extract	1 μL	1 μL	1 μL
ddH2O	to 10 μL	to 10 μL	to 10 μL
Reaction conditions
Incubation temperature	37 °C	37 °C	37 °C
Incubation time	15 minc–1 h	15 minc	1 h

^aThe volume of vector and insert DNAs can be adjusted. Ensure that the volume of unpurified DNA does not exceed 1/5 of total reaction volume

^bMolar ratio of insert to vector

^c15 min-incubation is enough for most SLiCE cloning tasks with a slightly reduced efficiency compared with 1 h