| 影响荧光终止法PCR循环测序反应的因素 |
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| 引用本文： | 钱锋,林汝仙,马淑华,王升启.影响荧光终止法PCR循环测序反应的因素[J].生物技术通讯,2000,11(3):206-208. |
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| 作者姓名： | 钱锋 林汝仙 马淑华 王升启 |
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| 作者单位： | 北京放射医学研究所,北京,100850 |
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| 摘 要： | 比较了一些影响荧光终止法ＰＣＲ循环测序反应的因素。实验结果显示在Ｂｅｃｋｍａｎ ＣＥＱ２０００自动测序仪上，可读序列长度随着ｐＵＣ１８模板量增加而逐渐增多，当模板量达到１２５ｎｇ时ＤＮＡ可读序列最长，以后随着ｐＵＣ１８量增加测序长度逐渐下降。当引物量是１μｌ时，其测序结果比用０．５μｌ引物时好。在同样模板量情况下，１０μｌ反应体积比５μｌ反应体积可读序列长。
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| 关 键 词： | 荧光终止法 PCR循环测序 模板 引物 |
Effects of concentrations of templates and primers on dye terminator cycle sequencing |
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| Qian Feng,Lin Ruxian,Ma Shuhua,Wang Shengqi.Effects of concentrations of templates and primers on dye terminator cycle sequencing[J].Letters in Biotechnology,2000,11(3):206-208. |
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| Authors: | Qian Feng Lin Ruxian Ma Shuhua Wang Shengqi |
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| Abstract: | Some parameters such as the amount of template and the amount of primer in Dye Terminator Cycle Sequencing (DTCS) are discussed in order to improve the sequencing quality in Beckman CEQ2000 automatic DNA sequencer. The most suitable amount of template in DTCS reaction is 125 nanogram; below or above 125 nanogram in template amount will result in low length readable DNA sequence. The sequencing results are better when using 1 micorliter primer than using 0.5 microliter. More length DNA sequences were read when DTCS was carried out in 10 microliter volume than in 5 microliter. |
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| Keywords: | dye terminator cycle sequencing template primer |
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