Abstract: In the process of constructing the tobacco DNA library, it often needs to process the massive data of DNA sequencing results, and carries on the sequence analysis and the management. According to the experience concrete request, the software for the management and analysis has been developed, it adopting .NET language, the new generation calculator language of the Microsoft Company, can easily read the sequencing report files, saving the result into the database automatically, and it has the function to distance Blast online. After the direct management and assistance analysis of more than one thousand sequences, new sequences of nucleic acid from tobacco has been found, then, we design the probe to carry on the relevant research of molecular biology by Tobacco Nucleic Acid Database Management System, and it enhances the efficiency of the entire experiment.