Uses the pCR™4Blunt-TOPO™ vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. Include the tools necessary to clone and select recombinant vectors containing your PCR fragment of choice. Resistant to Ampicillin (AmpR) and Kanamycin (KanR).
pCR4Blunt-TOPO Vector: Optimized for Sequencing
The pCR4Blunt-TOPO vector has a minimized multiple cloning site and, thus, a shorter distance between the sequencing primer sites and the insert site (as little as 33 bp). So sequencing reactions produce less vector sequence and more insert sequence. The pCR4Blunt TOPO vector has sites for four common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.
pCR4Blunt-TOPO Clone Selection and Manipulation
The pCR4Blunt-TOPO vector contains both ampicillin and kanamycin resistance markers and a LacZα-ccdB gene fusion for positive selection and blue/white screening. The vector's minimized multiple cloning site still includes flanking EcoRI sites for simplified excision of cloned PCR products and a unique Sse8387I site for generation of nested deletions prior to sequencing. T7 and T3 promoters are also present for in vitro transcription.
Simplified TOPO-based Cloning
Using TOPO cloning technology, there is no need for PCR primers containing specific sequences, post-PCR procedures, vector preparation, or other time-intensive DNA manipulation steps. Just add your PCR reaction straight to the provided topoisomerase-charged vector, incubate 5 minutes, and transform E. coli competent cells.
With up to 95% of clones carrying the desired insert, you can screen fewer clones, which saves time and money. The pCR4Blunt-TOPO vector has no overhangs for efficient ligation of PCR products created by proofreading thermostable polymerases that leave blunt-ended PCR products.
The Most Widely Used Cloning Kit
When it comes to cloning, TOPO cloning technology has been a reliable partner for thousands of scientists for over ten years. Fast, simple-to-use, and efficient, TOPO cloning has been applied to many different vectors for a wide array of applications.
Kit Options: Zero Blunt TOPO PCR Cloning Kits for Sequencing
Zero Blunt TOPO PCR Cloning kits for sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
- 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, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2895-20)
- Fast growth: Mach1™-T1R chemically competent E. coli (Cat. No. K2835-20)
- Provide your own cells (Cat. Nos. 450159 and 450031)
- Fast and easy: go from PCR to clones in just three steps, including a 5-minute cloning reaction
- Efficient: obtain up to 95% clones with correct insert
- Proven: enabling reliable performance for over a decade with over 20,000 citations
- Simple: no ligase, post-PCR procedures, or PCR primers containing specific sequences required
- Accurate: for use with high fidelity DNA polymerases, no need for A-overhang on PCR product
- Antibiotic resistance: Ampicillin (AmpR), Kanamycin (KanR)
Cloning, PCR Cloning
Shipping Condition: Dry Ice
|Kit, PCR Cloning|
|Cloning, PCR Cloning|
|Box 1: Topoisomerase I-activated pCR4Blunt-TOPO vector, Salt solution, dNTPs, Control template, T3, T7, M13 forward, and M13 reverse primers, Control PCR primers, Sterile water|