sentence: After the incubation , the cells were washed three times with hybridization buffer at 37 ° C , three times with 2xSSC at 37 ° C , once with 1xSSC at 37 ° C , once with 4xSSC at room temperature , once with 4xSSC containing 0.1 % Tween-20 at room temperature , and once with 4xSSC at room temperature .