주문정보
- - 재고수량은 변동될 수 있습니다.
- - 재고 확인 시 "갱신" 버튼을 누르시면 실시간 재고를 확인하실 수 있습니다.
- - 가격이 ‘별도문의‘ 시, 상단 ‘견적신청’ 버튼을 눌러 문의해주시면 빠른 답변을 받으실 수 있습니다.
| 선택 | Cat.No. | 제품명 | 가격(VAT별도) | 수량 |
|---|
제품특징
Product Name | Cat. No. | Size | Purification Kit Included | StellarCompetent Cells Included | CloneAmp HiFiPCR Premix Included |
In-Fusion HDCloning 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 HDCloning 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 oCloning 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 thereis a singlePCR product on the gel. oAlternatively, NucleoSpin Gel and PCR Clean-Up is suitable for gel extraction as well as PCR purification.
