怎样选择(设计测序用引物?

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  2019-09-12 10:24:58

  <p><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_18">测序用引物要求非常严格,不同于PR用引物。</span></p><p><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_19">(1)长度在15~25个碱基左右,一般选择20个碱基(根据GC含量作适当调整)</span></p><p><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_19"><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_19">(2)3&#39;端尽量选择G或C碱基</span></span></p><p></p><p><span class="result-text" style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;display: inline-block; vertical-align: top; color: rgb(51, 51, 51); width: 238px;Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_20">(3)Tm温度应选择50℃~70℃左右</span></p><p>(4)GC含量应选择在50%左右,尽量避开A、T、G、C的连续结构</p><p style=" font-family:&#39;Microsoft YaHei&#39;;font-size:16px;line-height: 2em;position: relative;">(5）避开引物自身形成发夹结构或引物二聚体结构等复杂结构</p><p style=" font-family:&#39;Microsoft YaHei&#39;;font-size:16px;line-height: 2em;position: relative;">(6）保证引物和模板匹配,特别是3端的几个碱基一定要匹配。同时必须严格保证引物和模板之间只能有一个结合位点</p><p><span class="result-text" style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;display: inline-block; vertical-align: top; color: rgb(51, 51, 51); width: 238px;Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_20"><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_21"></span></span><br/></p><p><span style="position:relative;line-height:2em;font-size:16px;font-family:Microsoft YaHei;color: rgb(51, 51, 51);Helvetica Neue&quot;, Helvetica, PingFangSC-Regular, &quot;Hiragino Sans GB&quot;, &quot;Microsoft YaHei&quot;, SimSun, sans-serif; background-color: rgb(250, 250, 250);" data-x-flag="10_19"></span><br/></p>

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