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| The Strategies of Obtaining Full-length Sequence by PCR Amplification Technology |
| LI Kun-peng1,2, ZHU Hua-bin2, HAO Hai-sheng2, ZHAO Xue-ming2, FENG Rong3, QIN Tong2, ZHANG Lin-bo1, WANG Dong2 |
1. College of Animal Science ＆Technology, Jilin Agriculture University, Changchun 130118, China;
2. The Key Laboratory for Farm Animal Genetic Resources and Utilization of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agriculture Sciences, Beijing 100193, China;
3. College of Wulanchabu Vocational, Jining 012000, China |
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Abstract Full-length sequence is the basic premise of the study on genetic function of new gene, three amplification methods of the full-length sequence of gene, inverse PCR, adapter-mediated PCR and random primer PCR, have been established based on PCR(polymerase chain reaction). Although the early established inverse PCR is inefficient in the ligation procedure and bring forth more non-specific amplification, it's extensively used in the identification of T-DNA insertion sites and gene fusion sites for its simplicity and quickness. With the improvement of the specificity, adapter-mediated PCR is getting more and more popular, many novel technologies, such as single specific primer PCR, capture PCR and step-down PCR, were derived, and the ideal amplification effects were obtained. Recently, owing to its high degree of automation, random primer PCR is playing an important role in the analysis of a large number of samples. With the discovery of powerful enzymes, such as, DNA polymerase, ligase and reverse transcriptase, the development of the strategies for obtaining full-length sequence by PCR amplification technology were promoted, many strategies were improved and integrated, and better experimental results were obtained, it provided more complete information for the study of gene structure, function and transcript characteristics.
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Received: 05 May 2012
Published: 25 November 2012
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