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用多级多聚酶链反应与液化低熔点琼脂糖凝胶中直接消化DNA相结合方法构建了8个突变体，经证实其重组DNA的阳性率为100％。从低熔点琼脂糖凝胶中切下目的DNA，在68℃完全熔化后用5倍体积的双蒸水稀释，该化合物即可直接用于酶解反应。混合物中凝胶比例以10g／L为宜。因而克服了凝胶中因目的DNA量少而纯化后回收率低无法进一步实验的弱点。
Eight mutants were constructed by multi-stage polymerase chain reaction combined with direct digestion of DNA in liquefied low-melting agarose gel. The positive rate of recombinant DNA was 100%. The target DNA was excised from the low melting point agarose gel, completely melted at 68 ° C and then diluted with 5 volumes of double distilled water. This compound was used directly in the enzymatic reaction. The gel ratio in the mixture is preferably 10 g / L. Thus overcoming the weakness that the gel can not be further experimentally recovered due to the low amount of target DNA and the low recovery after purification.