PCR引物在线设计软件的使用及操作流程,实例演示
1.The primary materials used in PCR(PCR反应所需的基本物质)
For a PCR reaction,four primary materials are needed, including dNTP--the building blocks for thenew DNA, template DNA--the DNA sequence that you want to amplify, DNApolymerase--a heat stable enzyme that catalyzes new DNA synthesis, Primers--single-strandedshort DNA (usually 16-50 nucleotides long) that are complementary to a shortregion on either end of the template DNA.The primers dictate the successfulnessof a PCR reaction.(在一个PCR反应中,需要四种基本物质,分别是dNTP,它是新DNA合成的原材料;模板DNA,即你所需扩增的目的DNA片段;DNA聚合酶,它是一种热稳定的酶能够催化新DNA的合成;以及引物,引物是一段较短的单链DNA,通常16-50个核苷酸的长度,能够与模板DNA上目的片段的任意一端互补配对。)
2.Some basic rules we need to keep in mind before designing.(引物设计的基本原则)
1).The primers should be closelycomplement with the target sequence.
第一,引物与模板的特定序列之间应紧密互补。
2). Avoid cross-dimer,self-dimerand hairpins.
第二,引物内部及引物之间避免形成二聚体或发夹结构。
3).Avoid non-specific amplification.
第三,避免非特异性扩增。