3.1. Steps involved in the construction of vectors using yeast
recombination-based cloning
Here, we provide a detailed ‘‘step-by-step’’ description of YRBC
and compare the procedure to the more conventional restriction/
ligation-based method (Fig. 1). Both methods require digestion of
the vector with suitable restriction enzymes. Conventional restriction/
ligation-based methods rely on the availability of suitable
restriction enzymes, which should generate the compatible ends
in both the vector and DNA fragment(s) of interest. By contrast,
YRBC requires linearization of the vector, but the enzyme can be
chosen freely and independently of the DNA fragment to be cloned
(henceforth named ‘‘insert’’). However, for both methods, the
digested vector needs to be purified.