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A number of primer design programs have been developed for diverse applications. However, none of these programs can be used to design primers for gene cloning aimed at expressing protein. Here we report the design of PrimerCE, which can be used to cover the whole process of gene cloning and expression. The main features of PrimerCE include inspection of restriction enzyme recognition sequence, open reading frame verification, stop codon inspection, base adjustment, primer optimization, sequence assembly and protein analysis. In addition to this, the program can be modified based on the different needs of users, e.g. new vector sequence and restriction enzyme recognition sequences can be integrated. With the use of PrimerCE, a pair of primers can be designed within minutes. The program has been proven to be efficient in designing primers in our high throughput cloning and gene expression experiments. The software is freely available at


Yinghao Cao, Jian Sun, Jianhui Zhu, Liyun Li, Guozhen Liu. PrimerCE: designing primers for cloning and gene expression. Molecular biotechnology. 2010 Oct;46(2):113-7

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PMID: 20373149

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