LR Reaction: Facilitates recombination of an attL substrate (entry clone) with an attR substrate (destination vector) to create an attB-containing expression clone (see the following diagram). This reaction is catalyzed by the Gateway®. LR Clonase® II enzyme mix.
Q. How long to incubate BP Clonase II in a microcentrifuge?
Q. What are ATT sites?
Site-specific integrative recombination of bacteriophage λ involves unequal partners. The minimal phage att site is composed of approximately 240-base pairs and four distinct binding sites for Int protein, at least three of which are crucial for function.
Q. What is Golden Gate reaction?
Golden Gate cloning is one of the easiest cloning methods in terms of hands-on time, as digestion and ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a single tube containing the Type IIS enzyme and ligase.
Table of Contents
Q. How long does it take to thaw Invitrogen BP Clonase II?
Thaw on ice the Invitrogen BP Clonase II enzyme mix for about 2 minutes. Vortex the BP Clonase II enzyme mix briefly twice (2 seconds each time). To each sample (Step 1, above), add 2 µl of BP Clonase II enzyme mix to the reaction and mix well by vortexing briefly twice.
Gateway® BP Clonase® II contains enzymes and buffer in a single mix to enable convenient ten-microliter reaction set up with fewer pipetting steps. For Research Use Only. Not for use in diagnostic procedures. Gateway™ BP Clonase™ II enzyme mix includes proteinase K solution (2 µg/µl) and a positive control vector. Store at -20°C or -80°C.
Q. What is the LR reaction in Clonase II?
Microcentrifuge briefly. Return BP Clonase II enzyme mix to –20°C or -80°C storage. Incubate reactions at 25°C for 1 hour. Add 1 µl of the Proteinase K solution to each sample to terminate the reaction. Vortex briefly. Incubate samples at 37°C for 10 minutes.
