Cycling parameters were 94 C for 2 min, then 35 cycles of
94 C for 10 s, 60 C for 20 s and 72 C for 2 min (30 s for 1 kb of
DNA), followed by a single 10 min extension at 72 C. The DNA
bands of interest were excised and purified from the gel as
described below. In parallel, the plasmid to be cloned was digested
with suitable restriction enzymes and the DNA fragment of interest
excised from the agarose ge