ADDITIONAL MATERIALS REQUIRED T4 DNA ligase
Taq DNA polymerase
Taq DNA polymerase buffer
TE buffer
CIAP
Falcon 2059 polypropylene tubes (15-ml)
Water baths (37°C and 42°C)
XL1-Blue Genotype: recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB lacIqZΔM15 Tn10 (Tetr)].
The XL1-Blue cells supplied with the pCMV-Tag vectors are not competent cells. Refer to Hanahan (1983) for a protocol for producing competent cells.