引物3'端单个错配对聚合酶链反应的影响。
Effect of single mismatches at 3'-end of primers on polymerase chain reaction.
作者信息
Simsek M, Adnan H
机构信息
Department of Biochemistry, Sultan Qaboos University, P.O.Box: 35, Postal Code: 123, Muscat, Sultanate of Oman.
出版信息
J Sci Res Med Sci. 2000 Jan;2(1):11-4.
Abstract
OBJECTIVE AND METHOD
To investigate the effect of three different mismatches (G/T, G/A or G/G) at the 3'-end of a primer to amplify a 268 bp (base pair) region of the human β-globin gene using different annealing temperatures (45 to 65°C).
RESULTS
The primer with the G/T mismatch was as efficient as the normal primer (G/C match) in the amplification of a 268 bp product at all temperatures tested. However, the primers having G/A or G/G mismatches at the 3'-end did not produce any specific polymerase chain reaction (PCR) fragment at all the annealing temperatures used, except a barely detectable 268 bp product for the G/G mismatch at 45 and 50°C.
CONCLUSION
We conclude that our PCR system was refractory to amplification when one of the primers contained a G/A or G/G mismatch at the 3'-end with template DNA.
摘要
目的与方法
使用不同退火温度（45至65°C），研究引物3'端三种不同错配（G/T、G/A或G/G）对扩增人β-珠蛋白基因268 bp（碱基对）区域的影响。
结果
在所有测试温度下，具有G/T错配的引物在扩增268 bp产物方面与正常引物（G/C匹配）效率相同。然而，3'端具有G/A或G/G错配的引物在所有使用的退火温度下均未产生任何特异性聚合酶链反应（PCR）片段，除了在45°C和50°C时，G/G错配产生了一个几乎检测不到的268 bp产物。
结论
我们得出结论，当其中一个引物在3'端与模板DNA存在G/A或G/G错配时，我们的PCR系统难以进行扩增。
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