Problems with and a system to eliminate single-primer PCR product contamination in simple sequence repeat molecular marker-assisted selection in soybean.
Problems with and a system to eliminate single-primer PCR product contamination in simple sequence repeat molecular marker-assisted selection in soybean.
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DOI：
10.4238/vol10-3gmr1366
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
J. Ma;S. Guan;D. Yao;Y. Wei;P. Wang
中科院分区：
文献类型：
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作者：
J. Ma;S. Guan;D. Yao;Y. Wei;P. Wang
聚合酶链反应（PCR）为大豆简单重复序列分子标记辅助选择（SSR MAS）提供了基础。该PCR体系及其各种条件已被许多研究人员优化。然而，目前对PCR体系优化的研究主要集中在双引物PCR产物上。通过对50份大豆样品进行单引物和双引物PCR产物的比较，发现在反应体系中使用单引物会导致部分SSR引物在PCR过程中扩增出片段，造成双引物PCR扩增的假阳性和片段不纯，不便于后续分析。采用“单引物PCR校正法”消除单引物非特异性PCR扩增的干扰，提高PCR质量。该方法可提高大豆SSR分子标记辅助选择的准确率和成功率。
Polymerase chain reaction (PCR) provides a foundation for simple sequence repeat molecular marker-assisted selection (SSR MAS) in soybean. This PCR system and its various conditions have been optimized by many researchers. However, current research on the optimization of the PCR system focuses on double-primer PCR products. We compared single- and double-SSR primer PCR products from 50 soybean samples and found that the use of single-PCR primers in the reaction system can lead to amplified fragments of portions of the SSR primers in the PCR process, resulting in both false-positives and fragment impurity of double-primer PCR amplification, inconvenient for subsequent analysis. We used "single-primer PCR correction" to eliminate interference caused by single-primer nonspecific PCR amplification and improve PCR quality. Using this method, the precision and success rates of SSR MAS in soybean can be increased.