IGEM:Peking/2007/Count:Ligation


 * Takara T4 DNA ligase
 * 1) Choose reaction volume: 10-20 μL
 * 2) Mix proper proportion (usually 1:1) of DNA fragment and vector.
 * 3) Add 10×(meaning 1/10 of final volume) ligase buffer.
 * 4) Add 1 μL ligase per 10 μL final volume.
 * 5) Bath overnight in 16℃ water, or longer in 4℃.
 * 6) Begin transformation.


 * New England Biolabs T4 DNA ligase
 * 1) Choose reaction volume: 10-20 μL
 * 2) Mix proper proportion (usually 1:1) of DNA fragment and vector.
 * 3) Add 10×(meaning 1/10 of final volume) ligase buffer.
 * 4) Add 0.5 μL ligase per 10 μL final volume.
 * 5) Bath in 16℃ water for 10 min, or in room temperature for convenience.
 * 6) Deactivation: 65℃ water for 10 min (longer ligation time or skipping deactivation step may lead to high pseudo-positive ratio.
 * 7) Begin transformation.