Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ MAX Efficiency DH5α-T1R E. coli
Zero Blunt&trade; TOPO&trade; PCR Cloning Kit, with One Shot&trade; MAX Efficiency DH5&alpha;-T1<sup>R</sup> <i>E. coli</i>
Invitrogen™

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ MAX Efficiency DH5α-T1R E. coli

Zero Blunt™ TOPO™ PCR Cloning Kits은 열안정성 polymerases 교정으로 증폭된 blunt-end PCR 산물 클로닝의 가장 신속하고 쉬운 고효율 방법을 제공합니다.자세히 알아보기
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카탈로그 번호수량
K282040
K2820-40으로도 사용됨
50 Reactions
K282020
K2820-20으로도 사용됨
25 Reactions
카탈로그 번호 K282040
K2820-40으로도 사용됨
제품 가격(KRW)
-
수량:
50 Reactions
Zero Blunt™ TOPO™ PCR Cloning Kits은 열안정성 polymerases 교정으로 증폭된 blunt-end PCR 산물 클로닝의 가장 신속하고 쉬운 고효율 방법을 제공합니다. 이 kit에는 linearized, topoisomerase I-activated pCR™-Blunt II-TOPO™ Vector가 들어있어 ligase없이 5분 내에 실험대에서 ligation이 가능합니다(1). pCR™-Blunt II-TOPO™ Vector의 기타 특징:

• ccdB 양성 선택 유전자(2)
• EcoR I 영역이 PCR 산물 insert 옆에 있어 insert를 쉽게 절개 가능함
• E. coli에서 선택한 Kanamycin 및 Zeocin™ 저항성 유전자
• SP6 promoter/primer site로 in vitro RNA transcription 및 sequencing 가능
• M13 forward/reverse primer site 로 sequencing 또는 PCR screening 가능

선택한 Zero Blunt™ TOPO™ PCR Cloning Kit는 PureLink™ Quick Plasmid Miniprep Kit (50 preps)과 함께 콤보 키트로도 제공되어 다음 단계 분석에서 TOPO™-cloned insert의 plasmid 정제를 신속하게 진행할 수 있습니다.
For Research Use Only. Not for use in diagnostic procedures.
사양
박테리아 또는 효모 균주DH5α
클로닝 방법Blunt TOPO™
제품라인One Shot
제품 유형PCR Cloning Kit
프로모터SP6
수량50 Reactions
벡터Blunt DNA Cloning Vectors
형식Kit
Unit SizeEach
구성 및 보관
Box 1:
• Linearized and topoisomerase I-activated pCR™Blunt II-TOPO™ vector
• Salt solution
• dNTPs
• Control template
• M13 forward and reverse primers
• Sterile water

Store at -5 to -30°C.
All reagents are stable for 6 months when properly stored.

Box 2:
• One Shot™ Chemically Competent or Electrocomp™ E. coli
• S.O.C. medium
• Supercoiled pUC19 control plasmid

Store at -68 to -85°C.

자주 묻는 질문(FAQ)

What is the difference between the pCR2.1-TOPO and pCR4-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCR4-TOPO vector has sequencing primer sites located as close as 33 base pairs from the PCR product insertion site. This minimizes the amount of vector DNA sequence that needs to be read before reaching the sequence of the insert, making the pCR4-TOPO vector very useful for sequencing applications.

What is the difference between the pCR2.1-TOPO and pCRII-TOPO vectors?

The vector backbones for both of these vectors are very similar. The main difference is that the pCRII-TOPO vector is a dual promoter vector, containing the SP6 and T7 promoters for in vitro transcription/sequencing, whereas the pCR2.1-TOPO vector contains only the T7 promoter for in vitro transcription/sequencing. Both vectors contain the M13 Forward and Reverse primer sites for sequencing or PCR screening.

Your TOPO cloning kits contain a control template and control primers. Can I obtain the sequence of the control template?

The sequence of the control template is proprietary.

What is the best molar ratio of PCR product:vector to use for TOPO TA cloning? Is there an equation to calculate the quantity to use?

We suggest starting with a molar ratio of 1:1 (insert:vector), with a range of 0.5:1 to 2:1. The quantity used in a TOPO cloning reaction is typically 5-10 ng of a 2 kb PCR product.

Equation:

length of insert (bp)/length of vector (bp) x ng of vector = ng of insert needed for 1:1 (insert:vector ratio)

What is the best ratio of insert:vector to use for cloning? Is there an equation to calculate this?

The optimal ratio is 1:1 insert to vector. Optimization can be done using a ratio of 0.5-2 molecules of insert for every molecule of the vector.

Equation:

length of insert (bp)/length of vector (bp) x ng of vector = ng of insert needed for 1:1 insert:vector ratio

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