Well-to-Well variation in qPCR amplification curves is suggestive of dysfunction in thermal cycler heat block

Modern quantitative polymerase chain reaction (qPCR) instrument typically has a 96 well heat block where the PCR tubes are placed for thermal cycling. With time and continued usage, there may be faster degradation of some wells relative to others, thereby, resulting in ineffective heating in some wells over others. This effect will manifest in different temperature cycling for different wells, which will likely result in different qPCR curves. Thus, observation of different qPCR amplification curves for different wells for the same experimental conditions is suggestive of dysfunction of the thermal heat block, or a more serious malfunction in the main system board that controls the temperature cycling of the heat block. This work presents a couple of representative amplification curves indicative of the effect.

现代实时定量聚合酶链式反应（quantitative polymerase chain reaction，qPCR）仪器通常搭载96孔热块，用于放置PCR反应管以开展热循环反应。随着使用时长增加与持续运转，部分孔的老化速率会快于其余孔，进而导致部分孔的加热效果劣于其他孔。该现象会表现为不同孔的热循环温度存在差异，最终可能引发qPCR扩增曲线出现不一致。因此，当同一实验条件下不同孔的qPCR扩增曲线出现差异时，提示该仪器的热块可能存在功能异常，或是控制热块热循环的主控电路板出现了更为严重的故障。本数据集提供了若干组可反映该故障效应的典型扩增曲线示例。