•     · 有效、可重复的多重突变分析
•     · 无需优化即可建立多重PCR反应
•     · 高特异性、灵敏的同时扩增所有片段
•     · 优化的操作流程，得到快速、可靠的结果
•     · 简单的操作指南，用于多种下游分析平台

Type-it Mutation Detect PCR Kit适用于快速、可靠的检测缺失、插入和异位等突变。该试剂盒基于高度特异的HotStarTaq Plus DNA Polymerase和获得专利的缓冲液系统，从而确保对突变位点进行可靠多重PCR扩增，无需优化。后续分析简单方便，可在琼脂糖凝胶、自动化电泳仪以及高分辨率毛细管测序仪上进行。

Type-it Mutation Detect PCR Kit优于其他供应商的试剂盒，确保可靠的突变分析。该试剂盒专门研发并经功能验证，可用于对缺失或插入等突变进行多重PCR分析，也可检测遗传修饰的有机体或微生物（参见" Sensitive detection of a mutated cancer-related gene）。该试剂盒也适合用作商业化SNP检测PCR体系的预扩增方法，如Applied Biosystems提供的SNaPshot体系（参见" Superior preamplification of SNPs"）。

Sensitive detection of a mutated cancer-related gene.

The indicated amounts of DNA extracted from a lymphoma-related cell line (Ramos) were spiked into human leukocyte DNA, as indicated in the figure, and the mutated Ramos target was detected along with 2 internal controls. Using the Type-it Mutation Detect Kit, the mutated 357 bp gene was detected even when only 25 pg of DNA (4 cells) was present. The lymphoma-relevant mutation could not be detected when using preoptimized conditions (Mg²⁺ and polymerase) with a hot-start DNA polymerase from Supplier I — even when using 25 ng of DNA (4000 cells). Electrophoresis was performed on a 1.3% agarose gel. M: 100 bp ladder.

Superior preamplification of SNPs.

Two different PCR systems (12-plex and 5-plex), both including the SNP of interest, were amplified using the standard protocol for the Type-it Mutation Detect PCR Kit (Q) or using preoptimized conditions with a hot-start DNA polymerase from Supplier A. With Type-it Mutation Detect Kits, all fragments of interest were amplified without any optimization of PCR parameters and could successfully be used as input for the SNaPshot PCR system. Even after optimization of Mg²⁺ concentration to 4 mM and varying the polymerase concentrations, amplification using the kit from Supplier A resulted in missing fragments. Electrophoresis was performed on the QIAxcel System. The first and the last bands on each lane are Upper Alignment Marker and Lower Alignment Marker, respectively. M: Alignment marker.

原理