Hébert B, Bergeron J, Potworowski E F, Tijssen P
Centre de recherche en immunologie, Institut Armand-Frappier, Laval, Québec, Canada.
Mol Cell Probes. 1993 Jun;7(3):249-52. doi: 10.1006/mcpr.1993.1036.
The use of mineral oil as a reaction mix overlay in conventional PCR may lead to problems. In addition to more difficult handling, traces of mineral oil in amplicon suspensions have been shown to decrease the efficiency of post-PCR manipulations. Commercial alternatives aimed at resolving the problem more than double the cost of an amplification. This is an important drawback when dealing with a large number of clinical samples. The use of inexpensive paraffin wax as a reaction mix overlay eliminated problems associated with the presence of mineral oil while being more practical and safer in handling potentially contaminated clinical samples. Moreover, when used in conjunction with a modified hot start technique, the use of paraffin wax increased the specificity and sensitivity of PCR amplifications over mineral oil in similar hot start conditions. Using the aforementioned method, the increase in specificity and sensitivity has enabled specific detection of viral DNA in clinical samples which the conventional PCR method failed to detect.
在传统PCR中使用矿物油作为反应混合物覆盖物可能会导致问题。除了操作更困难外，扩增子悬浮液中的微量矿物油已被证明会降低PCR后操作的效率。旨在解决该问题的商业替代品使扩增成本增加了一倍多。在处理大量临床样本时，这是一个重要的缺点。使用廉价的石蜡作为反应混合物覆盖物消除了与矿物油存在相关的问题，同时在处理潜在污染的临床样本时更实用、更安全。此外，当与改良的热启动技术结合使用时，在类似的热启动条件下，石蜡的使用比矿物油提高了PCR扩增的特异性和灵敏度。使用上述方法，特异性和灵敏度的提高使得能够在传统PCR方法未能检测到的临床样本中特异性检测病毒DNA。
