Invitrogen™

Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli

Catalog number: K2875J10
Invitrogen™

Zero Blunt™ TOPO™ PCR Cloning Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli

Catalog number: K2875J10
Catalog Number
K2875J10
also known as K2875-J10
Unit Size
10 reactions
Price (USD)
319.00
Your Price:
Availability
***
Quantity
Catalog Number
K287520
also known as K2875-20
Unit Size
25 reactions
Price (USD)
634.00
Your Price:
Availability
***
Quantity
Catalog Number
K287540
also known as K2875-40
Unit Size
50 reactions
Price (USD)
1,136.00
Your Price:
Availability
***
Quantity
Catalog NumberUnit SizePrice (USD)AvailabilityQuantity
K2875J10
also known as K2875-J10
10 reactions
319.00
Your Price:
***
K287520
also known as K2875-20
25 reactions
634.00
Your Price:
***
K287540
also known as K2875-40
50 reactions
1,136.00
Your Price:
***
Product Overview
Figures
Videos
Documents
FAQ
Citations & References
Additional Information
Recommendations
The Zero Blunt® TOPO® PCR Cloning Kit for Sequencing provides a highly efficient, 5-minute, one-step cloning strategy 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 (see figure) with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice.

pCR™4Blunt-TOPO® vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4Blunt-TOPO® vector to shorten the distance between sequencing primer sites and the insert site to as little as 33 bp. This means sequencing reactions give less vector sequence and more insert sequence. The pCR™4Blunt TOPO® vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4Blunt-TOPO® 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. 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 the provided E. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, which saves you time and money. The pCR™4Blunt-TOPO® vector used in this kit comes with no overhangs for efficient ligation of PCR products created by proofreading, thermostable polymerases that leave blunt-ended PCR products.

The standard in cloning
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.

Zero Blunt® TOPO® PCR Cloning Kit for Sequencing—kit formats
The Zero Blunt® TOPO® PCR Cloning Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending on what your needs are:

• General cloning: TOP10 cells (Cat. No. K2875-J10, K2875-20, K2875-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. 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)

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Bacterial or Yeast Strain
TOP10
Cloning Method
Blunt TOPO™
For Use With (Application)
Chromatin Biology
Product Line
One Shot™
Quantity
10 reactions
Vector
Blunt DNA Cloning Vectors
Product Type
PCR Cloning Kit
Promoter
T7, T3

Contents & Storage

Linearized and topoisomerase I-activated pCR™4-Blunt TOPO™ Vector, Salt Solution, dNTPs, Control Template and Primers, M13 Forward and Reverse Primers, One Shot™ Chemically Competent TOP10 E. coli, S.O.C. Medium, and a supercoiled control plasmid. Store One Shot™ E. coli at -80°C. Store all other components at -20°C. All reagents are guaranteed stable for 6 months when properly stored.

Figures

Frequently asked questions (FAQs)

Citations & References