LR Reaction using LR Clonase II

'' This is proportionally the same amount of enzyme per reaction volume, but it is a higher concentration of DNA than the Invitrogen protocol. ''

Materials

 * Destination Vector (~600 ng/µL)
 * Entry Clone (100-200 ng/µL)
 * LR Clonase II enzyme mix

Method

 * 1) Pipette 1 uL of Destination  Vector into a PCR tube.
 * 2) Add 1 uL of  Entry Clone  to the same tube.
 * 3) Finally, add 0.5 uL of LR Clonase II  to the mix.
 * 4) Gently tap the bottom of the tub to mix the reaction and incubate overnight at room temperature.   The LR reaction is catalyzed by incubating reactions at 25 C for 1 hour. Leaving the reaction ON results in higher efficiency and more transformants. Return LR Clonase II enzyme mix to -20 C or -80 C storage after use. 
 * 5) After overnight incubation, add 1 uL of Proteinase K solution to each sample to terminate the reaction. Vortex and centrifuge in a bench-top briefly. Incubate samples at 37 C for 10 minutes.
 * 6) Proceed with transformation.