Competitive PCR

Rapid determination of transgene copy number in tobacco by competitive PCR using a pair of SSR primers

K. Y. Xu, Wang, S. H., Xi, L., Wang, Q. J., Dong, C., Zhang, J. Y., Qu, S. C., and Zhang, Z., Rapid determination of transgene copy number in tobacco by competitive PCR using a pair of SSR primers, vol. 9, pp. 935-940, 2010.

We developed a straightforward, rapid, and inexpensive method to determine transgene copy number in tobacco. The plasmid (pSSRCopy) used for tobacco transformation contains a simple sequence repeat (SSR) locus, PT1199, which was partially deleted in the middle, a homogenous SSR locus in tobacco K326. A 168-bp segment of the cloned PT1199 was shortened to 95 bp by deleting a 73-bp internal fragment. Using a pair of SSR primers, competitive PCR was amplified from genomic DNA from transgenic tobacco harboring pSSRCopy, and the two expected bands were found.

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