分子克隆实验.pdf

pUC191.5kbDNApUC1301 pUC19 pUC1301 BamHI+KpnI pUC 1301 1.5kb pUC 19 r r amp kan α— pUC19 pUC1301 1 DNA DNA pU 1301 1.5kb pUC19 E.coil DNA T4DNA DNA TaqDNA T4RNA DNA CaCl2 DNA DNA LacZ IPTG X-gal X-gal 2 2.1 pUC19 DNA pU1301 2.2 2 Solution I: Tris.HCl pH 7.5 50 mM EDTA 10 mM RNase A 100 µg/ml Solution II: NaOH 0.2 M SDS 1 % Solution III: Final concentration KAC 1.32 M Using HAC to adjust pH to 4.8 TE (pH 8.0) Tris.HCl pH 8.0 10 mM EDTA pH 8.0 1 mM / 0.5 ×TBE 0.5 µg/ml EB 2.3 2.3.1 LA