Analysis of causes for formation of chimeras |
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Authors: | Xiaobo Zhang Xucheng Jin Hong Wang |
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Affiliation: | a Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Kaixuan Rd 268, Hangzhou 310029, China b Department of Biochemistry, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK, Canada S7N 5E5 |
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Abstract: | The present study investigated cDNA chimeras using two closely related members of the rice secretory protein gene family as an example. The chimeras detected in initial cDNA products that were amplified using LA Taq polymerase involved two categories: single-site type and multiple-site type with the frequency being about 20% and 3%, respectively. Further investigation revealed that PCR buffer additives and type of DNA polymerase had a major effect on the formation of chimeras in mixed-template amplification. Heteroduplex repair by microbial DNA repair systems in cDNA cloning was confirmed to produce the chimeras too, but it was not the major source. |
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