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Home > 전제품보기 > Cloning 관련 > In-Fusion Cloning > [NEW] In-Fusion® Snap Assembly
최고의 Colony 형성능과 정확한 PCR cloning

[NEW] In-Fusion® Snap Assembly

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제조사 제품코드 제품명 용량 가격
(부가세별도)
비고 사용자매뉴얼
Clontech
638947
In-Fusion® Snap Assembly Master Mix
관련학술 구매하기 라이선스 
10 회
281,000원  제조사 페이지로 바로가기 x111003, x111002
Clontech
638948
In-Fusion® Snap Assembly Master Mix
관련학술 구매하기 라이선스 
50 회
1,196,000원  제조사 페이지로 바로가기
Clontech
638949
In-Fusion® Snap Assembly Master Mix
관련학술 구매하기 라이선스 
250 회
3,889,000원  제조사 페이지로 바로가기


* 본 제품은 In-Fusion® HD Cloning kit (Code 639648)의 업그레이드 버전 제품입니다.

  • Subcloning 불필요 - Cloning 위치, insert 서열에 관계없이, 목적 vector에 바로 cloning
  • 높은 효율 - 0.5~15 kb의 다양한 크기의 fragment에서 95% 이상의 정확도
  • Seamless construction -불필요한 염기 추가 없이 cloning 완료 (e.g. 제한효소 사용 시, TA cloning 시)
  • 다양한 application - Single insert cloning은 물론 multiple cloning, mutagenesis 등 적용 가능 (더 많은 적용 사례를 보시려면 클릭하세요)

제품설명
In-Fusion® Snap Assembly는 15분만의 반응으로 선형화 vector에 어떤 insert라도 불필요한 염기 추가 없이 방향성 있는 cloning을 가능하다. Insert는 제한효소 처리나 phosphorylation, 혹은 blunt-end 처리 등 별도의 실험 과정이 필요하지 않다. In-Fusion® Snap Assembly의 효율은 95% 이상으로, 충분한 수의 colony를 얻을 수 있어 대량 분석을 위한 scale up에도 적용할 수 있다.
In-Fusion® Snap Assembly Master Mix은 ligase 없이도, insert와 선형화 vector 양 말단 15 bp의 상동 염기서열을 이용하여 효율적이고 정확하게 결합시킨다. 온라인에서 제공하는 In-Fusion® primer design tool을 이용하면, 각각의 실험에 알맞은 primer를 쉽게 디자인할 수 있다. 반응이 시작되면, In-Fusion® Master Mix는 선형화 된 DNA 가닥의 3’ end로부터 염기서열을 제거하고, 두 개의 DNA 단편에서 상보서열 간의 annealing을 통해 DNA가 결합시킨다. 이 방법은 Subcloning이나 별도의 조작 없이, 하나의 fragment뿐 아니라 여러 개의 fragment를 하나의 vector에 바로 적용할 수 있어 high throughput application에 적용할 수 있다.


그림 1. In-Fusion Snap Assembly와 In-Fusion HD (기존 제품) 간의 효율 비교
In-Fusion Snap Assembly는 기존 제품에 비해 정확도는 유지하면서, colony 형성 능력이 개선되었다.
Five different inserts (ranging in size from 405 bp to 1,005 bp) were simultaneously cloned into a 2.7-kb vector previously linearized by inverse PCR using In-Fusion HD and In-Fusion Snap Assembly. Each cloning reaction was performed in triplicate. Amplification primers and all reagents, except for the cloning master mixes, were the same between the two systems. After transformation and plating, 10 colonies from each replicate were analyzed by Sanger sequencing to determine the cloning accuracy. In-Fusion Snap Assembly yielded 4X more colonies than In-Fusion HD while maintaining high cloning accuracy.


그림 2. In-Fusion Snap Assembly와 NEBuilder HiFi (1)
PCR로 선형화한 vector에 single insert를 cloning한 결과, insert 크기에 관계없이 NEBuilder HiFi보다 In-Fusion Snap Assembly에서 훨씬 많은 colony 수를 보였다. 정확도는 95% 이상이었다.
A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.


그림 3. In-Fusion Snap Assembly와 NEBuilder HiFi (2)
제한효소로 선형화한 vector에 single insert를 cloning한 결과, 말단의 형태 (overhangs, blunt)에 관계없이 NEBuilder HiFi보다 In-Fusion Snap Assembly에서 훨씬 많은 colony 수를 보였다.
A single 3.8-kb insert was cloned into a standard cloning vector linearized by restriction digests to create 5' overhangs (Panel A), blunt ends (Panel B), or 3' overhangs (Panel C). These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers' specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing to determine the cloning accuracy. In-Fusion Snap Assembly yielded between 5X and 16X more colonies than NEBuilder HiFi, depending on the type of linearized vector ends.
Applications
- PCR cloning
- High-throughput cloning (see full range of reagent options on our high-throughput cloning page)
- Multiple-fragment cloning
- Gene synthesis
- Gene design
- Mutagenesis
- Domain swapping
- Domain modification
자세한 구성은 CoA를 참고하세요.
Citations
In-Fusion® HD Cloning kit 제품은 In-Fusion® Snap Assembly Master Mix로 업그레이드 되었습니다. In-Fusion® HD와 Citation이 동일합니다.

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