A reliable PCR amplification method for microdissected tumor cells obtained from paraffin-embedded tissue

Abebe Akalu , Juergen K.V Reichardt
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引用次数: 22

Abstract

We report a reliable method for PCR (polymerase chain reaction) amplification of genomic DNA from PET. This method uses DNA extraction with the QIAquick kit and amplification with AmpliTaq Gold. Amplification of up to 959 bp from PET was achieved with this combination which exceeds the current reported upper limit of 800 bp. In summary, the gradual activation of the AmpliTaq Gold during thermal cycling allows both for higher-fidelity and higher-throughput PCR amplification from PET. The use of the QIAquick kit for DNA purification of PET is sensitive, reproducible and suitable for management of a high number of samples.

从石蜡包埋组织中获得的微解剖肿瘤细胞的可靠PCR扩增方法
我们报告了一种可靠的PCR(聚合酶链反应)扩增PET基因组DNA的方法。本方法采用QIAquick试剂盒提取DNA, AmpliTaq Gold试剂盒扩增。该组合从PET中获得了高达959 bp的扩增,超过了目前报道的800 bp的上限。综上所述,在热循环过程中AmpliTaq Gold的逐渐激活允许PET进行更高保真度和更高通量的PCR扩增。使用QIAquick试剂盒对PET进行DNA纯化,灵敏度高,重复性好,适用于大量样品的管理。
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