Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments
<p>Abstract</p> <p>Background</p> <p>Multiplex PCR has been successfully applied in many areas since it was first reported in 1988; however, it suffers from poor universality.</p> <p>Results</p> <p>A novel method called Universal Multiplex PCR (U...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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BMC
2012-08-01
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| Series: | Plant Methods |
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| Online Access: | http://www.plantmethods.com/content/8/1/32 |
| _version_ | 1818759610739195904 |
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| author | Wen Daxing Zhang Chunqing |
| author_facet | Wen Daxing Zhang Chunqing |
| author_sort | Wen Daxing |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Multiplex PCR has been successfully applied in many areas since it was first reported in 1988; however, it suffers from poor universality.</p> <p>Results</p> <p>A novel method called Universal Multiplex PCR (UM-PCR) was created, which simultaneously amplifies multiple target fragments from genomic DNA. The method has two steps. First, the universal adapter-F and universal adapter-R are connected to the forward primers and the reverse primers, respectively. Hairpin structures and cross dimers of five pairs of adapter-primers are detected. Second, UM-PCR amplification is implemented using a novel PCR procedure termed “Two Rounds Mode” (three and 28–32 cycles). The first round (the first three cycles) is named the “One by One Annealing Round”. The second round (28–32 cycles) combines annealing with extension. In the first two cycles of the first round, primers only amplify the specific templates; there are no templates for the universal adapters. The templates of universal adapters begin to be synthesized from the second cycle of the first round, and universal adapters and primers commence full amplification from the third cycle of the first round.</p> <p>Conclusions</p> <p>UM-PCR greatly improves the universality of multiplex PCR. UM-PCR could rapidly detect the genetic purity of maize seeds. In addition, it could be applied in other areas, such as analysis of polymorphisms, quantitative assays and identifications of species.</p> |
| first_indexed | 2024-12-18T06:45:28Z |
| format | Article |
| id | doaj.art-54a7686429b645d4b54c5cd4684a6fac |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-18T06:45:28Z |
| publishDate | 2012-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-54a7686429b645d4b54c5cd4684a6fac2022-12-21T21:17:30ZengBMCPlant Methods1746-48112012-08-01813210.1186/1746-4811-8-32Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragmentsWen DaxingZhang Chunqing<p>Abstract</p> <p>Background</p> <p>Multiplex PCR has been successfully applied in many areas since it was first reported in 1988; however, it suffers from poor universality.</p> <p>Results</p> <p>A novel method called Universal Multiplex PCR (UM-PCR) was created, which simultaneously amplifies multiple target fragments from genomic DNA. The method has two steps. First, the universal adapter-F and universal adapter-R are connected to the forward primers and the reverse primers, respectively. Hairpin structures and cross dimers of five pairs of adapter-primers are detected. Second, UM-PCR amplification is implemented using a novel PCR procedure termed “Two Rounds Mode” (three and 28–32 cycles). The first round (the first three cycles) is named the “One by One Annealing Round”. The second round (28–32 cycles) combines annealing with extension. In the first two cycles of the first round, primers only amplify the specific templates; there are no templates for the universal adapters. The templates of universal adapters begin to be synthesized from the second cycle of the first round, and universal adapters and primers commence full amplification from the third cycle of the first round.</p> <p>Conclusions</p> <p>UM-PCR greatly improves the universality of multiplex PCR. UM-PCR could rapidly detect the genetic purity of maize seeds. In addition, it could be applied in other areas, such as analysis of polymorphisms, quantitative assays and identifications of species.</p>http://www.plantmethods.com/content/8/1/32Multiplex PCRPolymorphismsSpeciesGenetic purityMaize |
| spellingShingle | Wen Daxing Zhang Chunqing Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments Plant Methods Multiplex PCR Polymorphisms Species Genetic purity Maize |
| title | Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments |
| title_full | Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments |
| title_fullStr | Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments |
| title_full_unstemmed | Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments |
| title_short | Universal Multiplex PCR: a novel method of simultaneous amplification of multiple DNA fragments |
| title_sort | universal multiplex pcr a novel method of simultaneous amplification of multiple dna fragments |
| topic | Multiplex PCR Polymorphisms Species Genetic purity Maize |
| url | http://www.plantmethods.com/content/8/1/32 |
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