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GateV™ cloning and expression system utilizes the principle of specific recombination between λ phage and host Escherichia coli to effectively and quickly clone the target gene fragment into the starting vector to obtain the starting clone, and further quickly and accurately exchange and recombine from the starting clone to Different expression vectors were used to obtain expression clones. This method bypasses the cumbersome and inefficient classic enzyme-cut ligation method, thereby greatly simplifying and speeding up the construction process from a single gene to multiple expression vectors.
The GateV™ Cloning System consists of the following two multi-enzyme premix systems:
I. The GateV™ BP Master Mix is a multi-enzyme system that catalyzes the recombination between the target gene fragments (linear PCR fragments or fragments that have been cloned into vectors) containing attB sequences at both ends and the starting vectors containing attP sequences (such as pDONR™), resulting in the initial clone, also called the Entry clone, which contains attL sequences (sequences formed by the combination of attB and attP sequences after recombination) at both ends.
II. The GateV™ LR Master Mix is another multi-enzyme mixed system that catalyzes the recombination between the target gene containing attL sequences at both ends and the expression vector containing attR sequences, resulting in expression clones.
Its principle is shown in the figure below:
1. GateV™ BP Master Mix
Components |
Kit Size
|
|
C6001-10 (10 rxns) | C6001-20 (20 rxns) | |
BP Master Mix | 20 µl | 40 µl |
Proteinase K | 20 µl | 40 µl |
Control | 10 µl | 10 µl |
2. GateV™ LR Master Mix
Components |
Kit Size
|
|
C6002-10 (10 rxns) | C6002-10 (20 rxns) | |
LR Master Mix | 20 µl | 40 µl |
Proteinase K | 20 µl | 40 µl |
Control | 10 µl | 10 µl |
Epoch Biolabs (our old name)
Epoch Life Science (our correct name)
Epoch Life Sciences (typo by some authors)