用Primer Express软件来设计比较方便.docVIP

用Primer Express软件来设计比较方便、简单。网上搜索一下，我记得有网友提供的。如果实在找不到这个软件的话，可以参考下面的指南。先设计好Probe，再设计Primer。good luck。Design GuidelinesEnsure the following guidelines are met：? ?? ? × Amplicon length – 50 to 150 bases for optimum PCR efficiency.? ?? ? ×??Probe Length – 13 to 30 bases. Do not overlap primer and probe sequences.? ?? ? ×??Tm – 68 to 70 °C.? ?? ? ×??% GC – 30 to 80 %.? ?? ? ×??5 end – Cannot be a G residue. A G residue adjacent to the reporter dye will quench the reporter ? ?? ?? ?? ?fluorescence somewhat, even after cleavage.Avoid the following motifs：? ?? ? ×??Repeating oligonuc