KAPA高保真热启动酶(KAPAHiFi™ HotStart)是二代基因工程酶,保真性强,活力超高。基因工程改造后的KAPA高保真热启动酶，可以提高扩增产物的产量、酶的灵敏度、扩增速度、目标片段长度，扩增复杂模板的能力。

      一般的高保真酶扩增1kb碱基大约需要1-2分钟，而KAPA高保真热启动酶只需要30秒，可以节省75%的实验时间。KAPA高保真热启动酶具有很强的3’-5’外切酶，确保扩增准确性。KAPA高保真热启动酶，含有2种buffer，分别用于常规模板扩增和高难度模板扩增。

     1.超级保真;

     2.可以扩增各种复杂模板;

     3.扩增长度长,扩增基因组可超过15kb;

     4.扩增速度快,节省75%以上的时间。

     1.定点突变

     2.测序

     3.蛋白表达。

      图1.KAPA高保真酶扩增的出错率是野生型Taq酶的1/30，是Taq blends的1/10，是野生型Pfu的1/3。

     Figure 2. Amplification of hgDNA targets up to 11 kb. Each target was amplified from a descending range of template hgDNA concentrations (50 ng to 0.5 ng per reaction). Reactions (25 μl each) were performed using standard 3-step cycling profiles (35 cycles): 20 sec denaturation, 15 sec annealing, and 30 sec/kb extension time. Total reaction time for the 11 kb amplicon was 3h 50mins. 12.5 μl of each reaction was loaded on the gel.

     Figure3.  2 kb lambda phage target amplified from a 10 fold dilution series starting from 10 ng down to 10 fg using KAPAHiFi™ HotStart DNA Polymerase with Fidelity Buffer, fusion technology polymerase, and polymerase blend systems. All reactions (25 μl each) were performed using manufacturers’ protocols and buffers, with standard 3-step cycling profiles (35 cycles). 12.5 μl of each reaction was loaded on the gel.

 

     Figure 4. Amplification of AT- and GC-rich, single-copy human genomic targets. Seven single-copy gene fragments representing a range of GC content were used to compare the robustness of KAPAHiFi™ HotStart DNA Polymerase against a panel of competitor high fidelity DNA polymerases and polymerase blends. Amplicons range between 0.5 kb and 0.7 kb in length and ranged from 47% to 84% GC content. All reactions (25 μl each) contained 50 ng human genomic DNA as template and were performed using manufacturers’ protocols and buffers, with standard 3-step cycling profiles (35 cycles). 12.5 μl of each reaction was loaded on the gel.