Invitrogen™ Clonase™ Gateway™ LR Clonase II Enzyme Mix 11791020 Print Share Invitrogen™ Clonase™ Gateway™ LR Clonase II Enzyme Mix . Enzymes and buffer in single mix LR Clonase™ II enzyme mix is supplied as a 5X solution. 1. Add the following components to a 1.5 ml microcentrifuge tube at room temperature and mix: 1-3 µl of pENTR plasmid clone (50-150 ng total) 1 µl of pIGF-GTW destination vector (400 ng/µl) H2O to 4 µl. 2. LR Clonase® II Plus Enzyme Mix The LR Clonase® II Plus enzyme mix (Box 3) contains the followin g rea ents. Store Box 3 at -20°C for up to 6 months. For long-term storage, store at -80?C. Item Composition Quantity LR Clonase® II Plus Enzyme Mix Proprietary 40 ?l Proteinase K solution 2 ?g/?l in: 10 mM Tris-HCl, pH 7.5 20 mM CaCl 2 Gateway® LR Clonase™ II Enzyme Mix 20 reactions 11791-020 100 reactions 11791-100 Technology with Clonase™ II manual which is available for downloading from www.invitrogen.com or by contacting Technical Service (see page 35). New South Wales; Multibridge eclipse manual shifter; Wavefunction spartan 10 manual; Manual de sidunea en venezuela song; Atlas copco ga 160 vsd compressors manuals; Kenmore 15 10 the LR reaction that is catalyzed by the LR Clonase II enzyme mix that consists of integrase, integration host factor, and the phage excisionase (Fig. 1B). The LR clonase mix transfers the DNA fragment of interest ?ankedby two attL sites (in the entry clone) into a des-tination vector (pDEST) carrying two attR sites. After 4. To the remaining 10 ?L reaction, add: Destination vector (150 ng/?L) 2.0 ?L Gateway LR Clonase II enzyme mix 3.0 ?L Final volume 15 ?L 5. Mix well by vortexing briefly and incubate at 25°C for 2 hours. Note: Depending on your needs, the length of the recombination reaction can be extended up to 18 hours. 6. 5X LR Clonase™ reaction buffer 4 µl TE Buffer, pH 8.0 to 16 µl 2. Vortex LR Clonase™ enzyme mix briefly. Add 4 µl to the components above and mix well by vortexing briefly twice.