※ 본 제품은 종매되었습니다. 성능이 향상된 In-Fusion^® Snap Assembly 시스템으로 대체되었습니다.
※ In-Fusion Cloning 생생 후기보기

Product Name	Cat. No.	Size	Purification Kit Included	Stellar Competent Cells Included	CloneAmp HiFi PCR Premix Included
In-Fusion HD Cloning Plus	638909	10 rxns	Spin Columns	Yes	Yes
	638910	50 rxns	Spin Columns	Yes	Yes
	638911	100 rxns	Spin Columns	Yes	Yes
	638920	96 rxns	Spin Columns	Yes	Yes
In-Fusion HD Cloning Plus CE	638916	10 rxns	Cloning Enhancer	Yes	Yes
	638917	50 rxns	Cloning Enhancer	Yes	Yes
	638918	100 rxns	Cloning Enhancer	Yes	Yes
	638919	96rxns	Cloning Enhancer	Yes	Yes

Cloning Kit-PCR Based Seamless DNA Cloning with In-Fusion HD Cloning Plus

In-Fusion Cloning은 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 추가적은 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.

Subcloning 없이도 복수의 DNA 단편을 원하는 vector에 Cloning
내부의 특정 부분만 교체하는 Modular expression vector 제작
불필요한 염기가 없는 Fusion protein 구축
DNA 단편의 제거나 교체
Insert의 point mutation이나 제한효소 사이트 첨가
유전자내 형광 단백질의 tag 교체
cDNA로 UTRs첨가

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.

In-Fusion HD Cloning Plus Components

All In-Fusion HD Cloning Plus kits contain the In-Fusion HD enzyme premix and reagents for control experiments. In-Fusion HD Cloning Plus kits also include:

CloneAmp HiFi High Fidelity PCR Premix
CloneAmp HiFi PCR Premix is a convenient 2X master mix that provides exceptionally accurate, efficient and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid set-up of PCR reactions, and facilitates successful cloning.
Stellar Competent Cells
An E. coli HST08 strain is included that provides high transformation efficiency (greater than 5 x 10⁸ cfu/μg) and compliments the efficiency of the In-Fusion HD Cloning Plus kit.
Cloning Enhancer or NucleoSpin Gel and PCR Clean-Up
    o Cloning Enhancer (CE) is an easy-to-use enzyme premix that removes background plasmid DNA and PCR residue, eliminating
      the need for PCR insert purification prior to cloning. In-Fusion HD Cloning Plus CE kits are ideal for cloning when there is a single
      PCR product on the gel.
    o Alternatively, NucleoSpin Gel and PCR Clean-Up is suitable for gel extraction as well as PCR purification.