Figure 2

Verification of LIC adaptor sequence insertion into the pBABE-Neo plasmid. (A) PmlI enzyme-treated and untreated versions of both pBABE and pBLIC plasmids were run on a 0.7% agarose gel to check for insertion of the adaptor via presence of a PmlI site. A 1 kb ladder (NEB, N3232S) is used to mark band sizes. Gels were photographed on a Canon Powershot G10 mounted on a UVP Multidoc-it Digital Imaging System using standard preset contrast settings. Note that for the digested pBLIC, a single band corresponding to the linearized plasmid (~ 5 kb) is observed whereas the treated pBABE plasmid yields the same set of bands as the undigested plasmid. (B) Both pBABE and pBLIC were digested with SalI to check for loss of its restriction site, as expected upon successful insertion of the LIC adaptor sequence. For pBABE, a single band corresponding to a linearized plasmid at ~ 5 kb is seen whereas for pBLIC, the super-coiled and nicked forms are observed, indicating lack of digestion.