Thermo Fisher Scientific Zero Blunt TOPO PCR Cloning Kit for Sequencing, without competent cells, 25 Reactions

Invitrogen™ Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing (without competent cells)

Zero Blunt™ TOPO™ PCR Cloning kits for sequencing provide a highly efficient, 5-minute cloning reaction for the direct insertion of proofreading polymerase-amplified, blunt-ended PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4Blunt-TOPO™ vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include the tools necessary to clone and select recombinant vectors containing your PCR fragment of choice.

주요 특징

• Fast and easy — go from PCR to clones in just 3 steps, including a 5-minute cloning reaction
• Efficient — obtain up to 95% clones with correct insert
• Proven — reliable performance for over a decade with over 20,000 citations
• Simple — no ligase, post-PCR procedures, or specific sequence primers required
• Accurate — compatible with high-fidelity DNA polymerases; no need for A-overhang

pCR™4Blunt-TOPO™ Vector — Optimized for Sequencing

The vector has a minimized multiple cloning site and a shorter distance between sequencing primer sites and the insert site (as little as 33 bp), producing less vector sequence and more insert sequence.
It includes sites for four common sequencing primers: M13 forward, M13 reverse, T7, and T3, and aliquots of each are provided.

Clone Selection and Manipulation

The pCR™4Blunt-TOPO™ vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection.

• Flanking EcoRI sites for simplified excision of cloned PCR products
• Unique Sse8387I site for nested deletions prior to sequencing
• T7 and T3 promoters for in vitro transcription

Simplified TOPO™-based Cloning

Using TOPO™ cloning technology eliminates the need for PCR primers with specific sequences, post-PCR procedures, or vector preparation.
Simply add your PCR reaction to the topoisomerase-charged vector, incubate for 5 minutes, and transform E. coli competent cells.

Efficient Cloning

Up to 95% of clones carry the desired insert, reducing screening workload and saving time and cost. The vector’s blunt-end design supports efficient ligation of products generated by proofreading polymerases.

The Most Widely Used Cloning Kit

TOPO™ cloning technology has been trusted by scientists for over a decade, offering fast, simple, and efficient cloning for a wide range of applications.

Kit Options

Zero Blunt™ TOPO™ PCR Cloning kits for sequencing are available with various competent cells:

• General cloning: TOP10 cells (Cat. Nos. K2875-J10, K2875-20, K2875-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K2880-20, K2880-40)
• General cloning, T1 resistance: DH5α-T1R cells (Cat. No. K2895-20)
• Fast growth: Mach1™-T1R chemically competent E. coli (Cat. No. K2835-20)
• Provide your own cells (Cat. Nos. 450031, 450159)

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