※ In-Fusion Cloning 생생 후기보기

 *In-Fusion® HD Cloning Kit 는 In-Fusion® Snap Assembly Master Mix로 업그레이드 되었습니다.

PCR Cloning Kits―DNA Cloning―In Fusion HD Cloning System
In-Fusion Cloning System은 어떤 PCR 단편이나 복수의 DNA 단편도 선형화된 vector(linearized vector)에 15분(In-Fusion HD EcoDry는 30분)의 반응으로 directional cloning이 가능하다 (Figure 1). 또한 PCR 단편은 제한효소처리나 phosphorylation과 같은 추가적인 처리 단계가 필요하지 않다. In-Fusion 효소는 선형화된 vector 말단과 PCR로 증폭된 PCR 단편(insert)의 말단의 15bp homologue sequence를 인식하여 융합시킨다. 이 15 bp의 융합서열은 원하는 sequence를 증폭하고자 하는 PCR primer에 디자인하여 껴 넣으면 쉽게 만들 수 있다.

이 실험방법을 사용하면 아래와 같은 다양한 실험에 적용 가능하다.

Figure 1. The In-Fusion cloning protocol.
Figure 2. The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).


Figure 3. The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.
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