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All Answers2
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Geneing136
28 days ago
如果样品量小、主要做筛查且引物已验证，SYBR可控制成本。设计时避开二级结构和重复序列，单扩增子、短片段，复孔至少3次，并加入NRT、NTC、内参或外源对照。仪器要确认通道兼容，是否需要ROX校正；不同品牌Mix的荧光基线差异会影响阈值设置，切换时保留标准曲线。若病毒拷贝低、基质复杂或结果需对外报告，再上探针法。
余
余宇勇2259
27 days ago
低拷贝病毒核酸且要求高特异性时，优先考虑TaqMan：探针结合提高靶标识别，熔解曲线假峰风险低，适合单拷贝临界、标准曲线绝对定量、多靶标或合规场景。若预算紧，先做可行性验证：用已知梯度样本、强阳性弱阳性、NTC和引物二聚体对照，比较SYBR与探针法的检出下限和重复性。若SYBR出现非特异扩增或效率不稳，就不要为省钱强行替代。
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