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TOPO™ TA Cloning™ Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli (Supply Center Packaging) Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction, and are available with a variety of competent cells, or no competent cells, depending on your needs and budget. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice. This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

Features of TOPO® TA Cloning® Kits for Sequencing:

Get more sequence—allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations

TOPO® TA Cloning® Kits for Sequencing—overview

Vector: pCR™4-TOPO® TA vector—optimized cloning vector for improved sequencing results

Cloning method: TOPO® TA Cloning®—Topoisomerase I–based, 5-minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector

Competent cells: Various options—choose from kits with general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells, or use your own

pCR™4-TOPO® TA vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA 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™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA clone selection and manipulation
The pCR™4-TOPO® TA 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 with the providedE. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones and save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products, which contain 3´A overhangs.

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.

TOPO® TA Cloning® Kits for Sequencing—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)
• Provide your own: for flexibility and to save money (Cat. No. 450030)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TOPO™ TA Cloning™ Kit for Sequencing, with pCR™4-TOPO™ Vector, One Shot™ TOP10 Chemically Competent E. coli, and PureLink™ Quick Plasmid Miniprep Kit Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector 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.

Get More Sequence—Allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and Easy—Go from PCR-to-clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—Achieve up to 95% clones with correct insert
Proven—Reliable performance for over a decade with over 4000 citations

TOPO® TA Cloning® Kits for Sequencing Kit Overview
VECTOR: pCR™4-TOPO® TA Vector—Optimized cloning vector for improved sequencing results
CLONING METHOD: TOPO® TA Cloning —Topoisomerase I based 5 minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector
COMPETENT CELLS: Various Options —Choose from kits with either general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells

pCR™4-TOPO® TA VectorOptimized for Sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA 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™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA Clone Selection and Manipulation
The pCR™4-TOPO® TA 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 with the providedE. coli competent cells.

Efficient Cloning
With up to 95% of clones carrying the desired insert, you can screen less clones to help you save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products with 3´A overhangs.

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.

TOPO® TA Cloning® Kits for Sequencing Kit Options
The TOPO® TA 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 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

TOPO® products are For Research Use Only. Not intended for any animal or human therapeutic of diagnostic use.

Related Links
Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TOPO™ TA Cloning™ Kit for Sequencing, with One Shot™ TOP10 Chemically Competent E. coli Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector with specially designed sequencing primer sites that return more insert sequence and less vector sequence from each reaction, and are available with a variety of competent cells, or no competent cells, depending on your needs and budget. These kits include everything necessary to clone and select recombinant vectors containing your PCR fragment of choice. Features of TOPO® TA Cloning® Kits for Sequencing:

Get more sequence—allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations

TOPO® TA Cloning® Kits for Sequencing—overview

Vector: pCR™4-TOPO® TA vector—optimized cloning vector for improved sequencing results

Cloning method: TOPO® TA Cloning®—Topoisomerase I–based, 5-minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector

Competent cells: Various options—choose from kits with general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells, or use your own

pCR™4-TOPO® TA vectoroptimized for sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA 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™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA clone selection and manipulation
The pCR™4-TOPO® TA 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 with the providedE. coli competent cells.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones and save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products, which contain 3´A overhangs.

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.

TOPO® TA Cloning® Kits for Sequencing—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)
• Provide your own: for flexibility and to save money (Cat. No. 450030)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

Related Links

Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TOPO™ TA Cloning™ Kit, Dual Promoter, with pCR™II-TOPO™ Vector and One Shot™ TOP10 Electrocomp™ E. coli Invitrogen™

TOPO® TA Cloning® Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes (1). They use a pCR™-TOPO® Vector with covalently bound topoisomerase I for fast cloning and recombinants. pCR™-TOPO® Vectors include:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• Choice of T7 (pCR™2.1-TOPO®) or T7 and SP6 (pCR™II-TOPO®) promoters for in vitro RNA transcription and sequencing. Each vector also contains M13 forward and reverse primer sites for sequencing.
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and ampicillin resistance genes for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants

TOPO® TA Cloning® Kits are also available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

TOPO™ TA Cloning™ Kit, Dual Promoter, with One Shot™ TOP10 chemically competent E. coli cells Invitrogen™

TOPO® TA Cloning® Dual Promoter kits are for fast, efficient cloning and subsequent in vitro transcription. These kits include the pCR™II-TOPO® TA vector with dual T7 and SP6 promoters. By eliminating time-consuming and tedious restriction site cloning, TOPO® cloning is the most reliable cloning method, featuring a 3-step protocol and 5-minute cloning reaction, and yielding up to 95% recombinants.

Convenient features of this TOPO® TA Cloning® Dual Promoter kit include:
• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot® TOP10 Competent cells for added convenience and highest cloning efficiencies

Kit Options: TOPO® TA Cloning® Dual Promoter Kits
TOPO® TA Cloning® Dual Promoter kits can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:
• General cloning: TOP10 cells (Cat. Nos. K4600-J10, K4600-01, K4600-40)
• High-efficiency cloning: TOP10 Electrocomp™ cells (Cat. Nos. K4660-01, K4660-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. Nos. K4620-01, K4620-40)
• Fast growth: Mach1™ -T1R chemically competent E. coli (Cat. No. K4610-20)
• Repressor/induction needs: TOP10F’ cells (Cat. Nos. K4650-01, K4650-40)
• Provide your own cells (Cat. Nos. 451641, 450641)

TOPO™ TA Kit, TOP10F' (Supply Center Packaging) Invitrogen™

TOPO® TA Cloning® Kits for Subcloning provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® cloning') for the direct insertion of Taq polymerase–amplified PCR products into a plasmid vector for subcloning. Each kit uses the pCR™ 2.1-TOPO® TA vector with convenient restriction sites for subcloning. The cloning kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget. This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

Features of the TOPO® TA Cloning® Kits for Subcloning:

Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations
Simple—no ligase, post-PCR procedures, or PCR primers containing specific sequences are required

TOPO® TA Cloning® Kits for Subcloning—overview

Vector: pCR™ 2.1-TOPO® TA vector—subcloning vector with 15 convenient restriction sites flanking your insert for easy, directional subcloning
Cloning method: TOPO® TA Cloning® —Topoisomerase I–based, 5-minute ligation of PCR products with 3′-A overhangs (Taq polymerase amplified) to TOPO® vector with T overhangs
Competent cells: various options—choose from kits with either general, high-efficiency, bacteriophage T1–resistant, fast-growing competent cells, or use your own

pCR™ 2.1-TOPO® TA vector—cloning and subcloning simplicity
The pCR™ 2.1-TOPO® TA vector is linearized with 3'-thymidine (T) overhangs for direct ligation of Taq polymerase–amplified PCR products (TA cloning®) and is 'activated' with covalently bound Topoisomerase I. There is no need to add ligase and cloning is complete in 5 minutes. EcoRI sites flanking the PCR product insertion site allow for easy excision of inserts or use any combination of 15 convenient restriction sites flanking your PCR insert for easy, directional subcloning.

pCR™ 2.1-TOPO® TA clone selection and manipulation
The pCR™ 2.1-TOPO® TA vector contains both ampicillin and kanamycin resistance markers and the LacZα gene for blue/white screening.

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.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones, saving time and money. The pCR™ 2.1-TOPO® TA vector used in this kit comes with 3'-T overhangs for efficient ligation of Taq polymerase–amplified PCR products, which contain 3'-A overhangs.

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.

TOPO® TA Cloning® Kits for Subcloning—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 cells (Cat. No. K4500-01, K4500-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4560-01, K4560-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. No. K4520-01, K4520-40)
• Fast growth: Mach1™ -T1R Chemically Competent E. Coli (Cat. No. K4510-20)
• Provide your own: for flexibility and to save money (Cat. No. 450641)

We also offer two versions of the kit that include a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4500-02 and K4510-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

TA Cloning™ Kit, with pCR™2.1 Vector, without competent cells Invitrogen™

The TA Cloning® Kit with pCR™2.1 vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The TA Cloning® Kit uses the pCR™2.1 cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.

Features of the TA Cloning® Kit with pCR™2.1 vector:
Fast & convenient—15-minute, room-temperature ligation
Efficient—blue/white screening and >80% clones with correct insert
Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
Hassle-free—eliminates any enzymatic modifications of the PCR product
Streamlined—does not require the use of PCR primers that contain restriction sites

The pCR™2.1 vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 promoter for in vitro RNA transcription and sequencing
• A versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

How TA Cloning® Works
Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Kit Configurations
The TA Cloning® Kit is offered in a variety of configurations: without competent cells (K2020-20 and K2020-40), with One Shot® INVF' Chemically Competent E. coli (K2000-01 and K2000-40), with One Shot® TOP10F' Chemically Competent E. coli (K2030-01 and K2030-40), and with One Shot® TOP10 Chemically Competent E. coli (K2040-01 and K2040-40) in 20- and 40-reaction kit sizes.

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

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.

• 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—enabling reliable performance for over a decade with over 20,000 citations
• Simple—no ligase, post-PCR procedures, or PCR primers containing specific sequences are required
• Accurate—for use with high fidelity DNA polymerases, no need for A-overhang on PCR product

pCR™4Blunt-TOPO® Vector—Optimized for Sequencing
The pCR™4Blunt-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 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 E. coli competent cells.

Efficient Cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, which helps save time and money. The pCR™4Blunt-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 Kit 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. 450031, 450159)

TOPO™ TA, Dual TOP10 (Supply Center Packaging) Invitrogen™

The TOPO® TA Cloning® Dual Promoter Kit is for fast, efficient cloning and subsequent in vitro transcription/translation. This kit includes the pCR™II-TOPO® TA vector (see figure). By eliminating time-consuming and tedious restriction site cloning, TOPO® cloning is the most reliable method, taking only 5 minutes using a 3-step protocol, and yielding up to 95% recombinants. Convenient features of the TOPO® TA Cloning® Dual Promoter Kit include:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and SP6 promoters for efficient in vitro transcription
• M13 forward and reverse primer sites for sequencing
• 16 convenient restriction sites, including EcoRI, flanking your insert for subsequent excision or subcloning
• Kanamycin and ampicillin resistance for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants
• Includes OneShot® TOP10 Competent cells for added convenience and highest cloning efficiencies

This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

TOPO™ TA Cloning™ Kit, with One Shot™ MAX Efficiency™ DH5α-T1R E. coli Invitrogen™

TOPO® TA Cloning® Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes (1). They use a pCR™-TOPO® Vector with covalently bound topoisomerase I for fast cloning and recombinants. pCR™-TOPO® Vectors include:

• 3´-T overhangs for direct ligation of Taq-amplified PCR products
• Choice of T7 (pCR™2.1-TOPO®) or T7 and SP6 (pCR™II-TOPO®) promoters for in vitro RNA transcription and sequencing. Each vector also contains M13 forward and reverse primer sites for sequencing.
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and ampicillin resistance genes for your choice of selection in E. coli
• Easy blue/white colony screening for selection of recombinants

TOPO® TA Cloning® Kits are also available in combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.

TOPO™ TA Cloning™ Kit for Sequencing, with One Shot™ TOP10 Electrocomp™ E. coli Invitrogen™

The TOPO® TA Cloning® Kits for Sequencing provide a highly efficient, 5 minute, one-step cloning strategy ('TOPO® Cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for sequencing. Each kit uses the pCR™4-TOPO® TA vector 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.

Get More Sequence—Allows for more insert sequence and less vector seuquence when using standard sequencing primers
Fast and Easy—Go from PCR-to-clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—Achieve up to 95% clones with correct insert
Proven—Reliable performance for over a decade with over 4000 citations

TOPO® TA Cloning® Kits for Sequencing Kit Overview
VECTOR: pCR™4-TOPO® TA Vector—Optimized cloning vector for improved sequencing results
CLONING METHOD: TOPO® TA Cloning —Topoisomerase I based 5 minute ligation of PCR products with 3´A overhangs (Taq-amplified) to the vector
COMPETENT CELLS: Various Options —Choose from kits with either general, high-efficiency, bacteriophage T1-resistant, or fast-growing competent cells

pCR™4-TOPO® TA VectorOptimized for Sequencing
We have removed much of the multiple cloning site from the pCR™4-TOPO® TA 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™4-TOPO® TA vector has sites for 4 common sequencing primers: M13 forward, M13 reverse, T7, and T3. The kits include an aliquot of each.

pCR™4-TOPO® TA Clone Selection and Manipulation
The pCR™4-TOPO® TA 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 with the providedE. coli competent cells.

Efficient Cloning
With up to 95% of clones carrying the desired insert, you can screen less clones to help you save time and money. The pCR™4-TOPO® TA vector used in this kit comes with 3´T overhangs for efficient ligation of Taq-amplified PCR products with 3´A overhangs.

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.

TOPO® TA Cloning® Kits for Sequencing Kit Options
The TOPO® TA 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 (Cat. No. K4575-J10, K4575-01, K4575-40)
• High efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4580-01, K4580-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K4595-01, K4595-40)
• Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K4530-20)

We also offer a version of the kit that includes a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4575-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

TOPO® products are For Research Use Only. Not intended for any animal or human therapeutic of diagnostic use.

Related Links
Custom Vector Construction and Cloning Services
Plasmid DNA Purification Kit Selection Guide
PCR Reagents, Instruments, and Supplies

TA Cloning™ Kit, Dual Promoter, with pCR™II Vector and One Shot™ INVαF' Chemically Competent E. coli Invitrogen™

The TA Cloning® Kit Dual Promoter with pCR™II vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The T7 and Sp6 promoters of the pCR™II vector allow in vitro transcription of the insert to produce sense or anti-sense products. The TA Cloning Kit Dual Promoter uses the pCR™II cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.

Features of the TA Cloning® Kit Dual Promoter:
Fast & convenient—15-minute, room-temperature ligation
Efficient—blue/white screening and >80% clones with correct insert
Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
Hassle-free—eliminates any enzymatic modifications of the PCR product
Streamlined—does not require the use of PCR primers that contain restriction sites

The pCR™II vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 and Sp6 promoters for in vitro RNA transcription and sequencing
• Versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

How TA Cloning® Works
Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Kit Configurations
The TA Cloning® Kit is offered in a variety of configurations: with One Shot® INVF' Chemically Competent E. coli (K2050-01 and K2050-40), One Shot® TOP10F' Chemically Competent E. coli (K2060-01 and K2060-40), and without competent cells (K2750-20 and K2750-40) in 20- and 40-reaction kit sizes.

TOPO™ TA Cloning™ Kit for Subcloning, with TOP10F' E. coli Invitrogen™

TOPO® TA Cloning® Kits for Subcloning provide a highly efficient, 5-minute, one-step cloning strategy ('TOPO® cloning') for the direct insertion of Taq polymerase-amplified PCR products into a plasmid vector for subcloning. Each kit uses the pCR™ 2.1-TOPO® TA vector with convenient restriction sites for subcloning. The cloning kits are available with a variety of competent cells, or without competent cells, depending upon your needs and budget. Features of the TOPO® TA Cloning® Kits for Subcloning:

Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations
Simple—no ligase, post-PCR procedures, or PCR primers containing specific sequences are required

TOPO® TA Cloning® Kits for Subcloning—overview

Vector: pCR™ 2.1-TOPO® TA vector—subcloning vector with 15 convenient restriction sites flanking your insert for easy, directional subcloning

Cloning method: TOPO® TA Cloning® —Topoisomerase I-based, 5-minute ligation of PCR products with 3′-A overhangs (Taq polymerase amplified) to TOPO® vector with T overhangs

Competent cells: various options—choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells, or use your own

pCR™ 2.1-TOPO® TA vector—cloning and subcloning simplicity
The pCR™ 2.1-TOPO® TA vector is linearized with 3'-thymidine (T) overhangs for direct ligation of Taq polymerase-amplified PCR products (TA cloning®) and is 'activated' with covalently bound Topoisomerase I. There is no need to add ligase and cloning is complete in 5 minutes. EcoRI sites flanking the PCR product insertion site allow for easy excision of inserts or use any combination of 15 convenient restriction sites flanking your PCR insert for easy, directional subcloning.

pCR™ 2.1-TOPO® TA clone selection and manipulation
The pCR™ 2.1-TOPO® TA vector contains both ampicillin and kanamycin resistance markers and the LacZα gene for blue/white screening.

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.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen fewer clones, saving time and money. The pCR™ 2.1-TOPO® TA vector used in this kit comes with 3'-T overhangs for efficient ligation of Taq polymerase-amplified PCR products, which contain 3'-A overhangs.

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.

TOPO® TA Cloning® Kits for Subcloning—kit options
The TOPO® TA Cloning® Kit for Sequencing can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

• General cloning: TOP10 cells (Cat. No. K4500-01, K4500-40)
• High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K4560-01, K4560-40)
• General cloning, bacteriophage T1 resistance: DH5α-T1R cells (Cat. No. K4520-01, K4520-40)
• Fast growth: Mach1™ -T1R Chemically Competent E. Coli (Cat. No. K4510-20)
• Provide your own: for flexibility and to save money (Cat. No. 450641)

We also offer two versions of the kit that include a PureLink™ Quick Plasmid Miniprep Kit (Cat. No. K4500-02 and K4510-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.

TA Cloning™ Kit, with pCR™2.1 Vector and One Shot™ TOP10F' Chemically Competent E. coli Invitrogen™

The TA Cloning® Kit with pCR™2.1 vector provides a quick, one-step cloning strategy for directly inserting a Taq-amplified PCR product into a plasmid vector. The TA Cloning® Kit uses the pCR™2.1 cloning vector and ExpressLink™ T4 DNA Ligase to generate a ligation product in a fifteen-minute, room-temperature ligation step. Reactions typically yield >80% recombinants containing inserts.

Features of the TA Cloning® Kit with pCR™2.1 vector:
Fast & convenient—15-minute, room-temperature ligation
Efficient—blue/white screening and >80% clones with correct insert
Flexible—choice of kanamycin or ampicillin resistance for flexible antibiotic selection
Hassle-free—eliminates any enzymatic modifications of the PCR product
Streamlined—does not require the use of PCR primers that contain restriction sites

The pCR™2.1 vector provides:
• 3'-T overhangs for direct ligation of Taq-amplified PCR products
• T7 promoter for in vitro RNA transcription and sequencing
• A versatile polylinker with flanking EcoR I sites for easy excision of inserts
• M13 forward and reverse primer sites for sequencing

How TA Cloning® Works
Taq polymerase has a non-template-dependent activity that adds a single deoxyadenosine (A) to the 3' ends of PCR products. The linearized vector supplied in this kit has single 3' deoxythymidine (T) residues. This allows PCR inserts to ligate efficiently with the vector.

Kit Configurations
The TA Cloning® Kit is offered in a variety of configurations: without competent cells (K2020-20 and K2020-40), with One Shot® INVF' Chemically Competent E. coli (K2000-01 and K2000-40), with One Shot® TOP10F' Chemically Competent E. coli (K2030-01 and K2030-40), and with One Shot® TOP10 Chemically Competent E. coli (K2040-01 and K2040-40) in 20- and 40-reaction kit sizes.

Zero Blunt™ TOPO™ PCR Cloning Kit, with One Shot™ TOP10 Chemically Competent E. coli (Supply Center Packaging) Invitrogen™

Zero Blunt® TOPO® PCR Cloning Kit provides a highly efficient, 5-minute, one-step cloning strategy ("TOPO® Cloning") for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. Each kit has the Zero Blunt® TOPO® vector (see figure) containing the ccdB gene for positive selection, only permitting growth of plasmid vectors with recombinants. Zero® Blunt® TOPO® kits are available with a variety of competent cells, or without competent cells, depending on your needs and budget. This product comes in Supply Center packaging, which features an additional outer box that holds both the TOPO® vector box and the competent cells box. This special packaging helps ensure that visitors to a Supply Center obtain the complete kit.

Features of the Zero® Blunt® TOPO® PCR Cloning Kits:

Fast and easy—go from PCR to clones in just 3 steps and in as little as 5 minutes hands-on time
Efficient—obtain up to 95% clones with correct insert
Proven—reliable performance for over a decade with over 4,000 citations
Simple —no ligase, post-PCR procedures, or PCR primers containing specific sequences are required.

Zero Blunt® TOPO® Cloning Kits—overview

Vector: pCR™Blunt II-TOPO® vector—contains the ccdB gene, allowing for direct selection of recombinants

Cloning method: TOPO® cloning—Topoisomerase I–based, 5-minute ligation of blunt-ended PCR products amplified with proofreading thermostable polymerases

Competent cells: Various options—choose from kits with either general, high-efficiency, bacteriophage T1-resistant, fast-growing competent cells, or use your own.

pCR™Blunt II-TOPO® vector—designed for proofreading polymerases
The pCR™Blunt II-TOPO® vector is designed to clone blunt-ended PCR products generated by thermostable proofreading polymerases such as Platinum® Pfx DNA Polymerase. The pCR™Blunt II-TOPO® vector contains:

• EcoRI sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and Zeocin™ resistance genes for your choice of selection in E. coli
• SP6 promoter/primer site for in vitro RNA transcription and sequencing
• M13 forward and reverse primer sites for sequencing or PCR screening

pCR™Blunt II-TOPO® clone selection
pCR™-Blunt II-TOPO® allows for direct selection of recombinants via disruption of the lethal E. coli gene, ccdB. The vector contains the ccdB gene fused to the C-terminus of the LacZα fragment. Ligation of a blunt-end PCR product disrupts expression of the LacZα-ccdB gene fusion, permitting growth of only positive recombinants upon transformation. Cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white screening is not required.

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.

Efficient cloning
With up to 95% of clones carrying the desired insert, you can screen less clones, saving you time and money. The pCR™Blunt II-TOPO® vector used in this kit comes with lethal ccdB gene for efficient selection of blunt-ended PCR recombinants products generated bythermostable proofreading polymerases.

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 options
The Zero Blunt® TOPO® PCR Cloning Kit for direct insertion of blunt-ended PCR products into a plasmid vector can be purchased with a variety of competent cells that deliver different advantages depending upon your needs:

General cloning: TOP10 cells (Cat. No. K2800-20, K2800-40)
High-efficiency cloning: TOP10 Electrocomp™ Cells (Cat. No. K2860-01, K2860-40)
General cloning, bacteriophage T1 resistance: DH5α-T1R (Cat. No. K2820-20, K2820-40)
Fast growth: Mach1™-T1R Chemically Competent E. Coli (Cat. No. K2830-20)
Provide your own: for flexibility and to save money (Cat. No. 450254)

We also offer a version of the kit that includes a PureLink® Quick Plasmid Miniprep Kit (Cat. No. K2800-02) for use in isolation of clean, sequencing-ready, recombinant plasmid.
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