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Evaluate the Efficiency of AGILENT SureDIrect Blood PCR Kit

机译：评估安捷伦患者血液PCR套件的效率

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In medical fields, the interest for direct amplification from blood has been increased rapidly, leading to the growing demand for direct DNA amplification from blood However, this direct assay may encounter major obstacle from potential inhibitors that present as blood components, or be introduced during preservation such as anticoagulants. Among commercial direct PCR kits, recently, the AGILENT SureDirect Blood PCR Kit had been developed and recommended as a convenient tool with powerful inhibitor tolerance for rapid PCR directly on blood samples. Accordingly, the aim of this study is to obtain optimal conditions and evaluate the efficiency of direct PCR by the SureDirect Kit in used for genetic diagnosis purpose. Target to be amplified was a highly conserved 121-bp sequence located on miRNA196a2. The archived optimal conditions for the SureDirect Kit, 5% (vol/vol) of whole blood sample and l0μL final volume, were validated successfully on 44 EDTA-blood samples frozen for 1 year and 29 EDTA-blood samples in 2 conditions: fresh and 1-month freezing. None of validation reactions were failed, making the successful rate of the SureDirect Kit, when perform with the archived optimal conditions, approach to 1.0. Besides, it was suggested that storage duration seems to have subtle effect on lowering PCR product quality but no significant effect on the success of direct amplification. In conclusion, the AGILENT SureDirect Blood PCR Kit is suggested as an applicable and economical direct PCR tool for genetic diagnosis purposes.
机译：在医疗领域，从血液直接扩增的兴趣已经迅速增加，导致对血液的直接DNA扩增的需求不断增长，这种直接测定可能会遇到作为血液成分的潜在抑制剂的主要障碍，或者在保存期间引入如抗凝血剂。在商业直接PCR套件中，最近，已开发安置安捷伦Syedirect血液PCR套件，并推荐作为一种方便的工具，其具有直接在血样上快速PCR的抑制剂耐受性。因此，本研究的目的是获得最佳条件，并评估SUREDIRECT试剂盒用于遗传诊断目的的直接PCR的效率。要扩增的靶标是位于miRNA196a2上的高度保守的121​​bp序列。 SurediRect试剂盒，全血样品和L0μl最终体积的存档最佳条件，在24个EDTA-血液样品中成功验证了1年和29种EDTA-血液样品的29个条件：新鲜和1个月冻结。没有一项验证反应失败，既有成功率，何时服用存档的最佳条件，接近1.0。此外，有人建议储存持续时间似乎对降低PCR产品质量但对直接扩增成功没有显着影响。总之，Agilent Suredirect血液PCR套件被建议作为遗传诊断目的的适用和经济的直接PCR工具。

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