Of particular importance and interest are the effects of pulsed Nd:YAG laser beam welding parameters on penetration and microstructure characterization of DP1000 butt joint, which is widely used in the automotive industry nowadays. Some key experimental technologies including pre-welding sample preparation and optimization design of sample fixture for a sufficient shielding gas flow are performed to ensure consistent and stable testing. The weld quality can be influenced by several process factors, such as laser beam power, pulse duration, overlap, spot diameter, pulse type, and welding velocity. The results indicate that these key process parameters have a significant effect on the weld penetration. Meanwhile, the fusion zone of butt joints exhibits obviously greater hardness than the base metal and heat affected zone of butt joints. Additionally, the volume fraction of martensite of dual-phase steel plays a considerable effect on the hardness and the change of microstructure characterization of the weld joint.