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TOPO® Cloning Technology
Fast, effective cloning of PCR products

Direct your cloning future.
TOPO® Cloning Technology

With TOPO® Cloning
Technology You Can:
• Clone Taq-amplified,
blunt-end, and long
PCR products
• Sequence or clone
directly into an
expression vector
• Ligate in 5 minutes at
room temperature and
obtain up to 99%recombinants
These products may be covered by one or more Limited Use Label Licenses (See the Invitrogen catalog or our web site,
www.invitrogen.com). By the use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses.
For research use only. Not intended for any animal or human therapeutic or diagnostic use. Printed in the U.S.A. ©2003 Invitrogen
Corporation.All rights reserved. Reproduction forbidden without permission.

Corporate headquarters:
1600 Faraday Avenue • Carlsbad, CA 92008 USA • Tel: 760 603 7200 • Fax: 760 602 6500 • Toll Free Tel: 800 955 6288 • E-mail: tech_service@invitrogen.com • www.invitrogen.com

European headquarters:
Invitrogen Ltd 3 • Inchinnan Business Park • Fountain Drive • Paisley PA4 9RF, UK • Tel: +44 (0) 1 81 6100• Fax: +44 (0) 1 81 6260 • E-mail: eurotech@invitrogen.com
41 4
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710-021849 070803

10M

• Obtain up to 99% recombinants
• Ligate in 5 minutes at room temperature
• Clone Taq-amplified, blunt-end, and long PCR products

With TOPO® Cloning Kits You Can:

TOPO® Cloning Kits

Powerful PCR Cloning Tools
For optimal cloning results, you need a technology that you
can rely on.TOPO® Cloning is the most effective technology
available for cloning PCR products and other DNA molecules.
It yields up to 99% recombinants via a simple 5-minute,
bench-top ligation.

Topoisomerase greatly improves cloning
TOPO® Cloning makes ligation faster and more successful.

3´ thymidine. It cleaves one DNA strand, enabling the

It enables 5-minute, bench-top ligation and yields up toDNA to unwind. The enzyme then religates the ends of

99% recombinants. This speed and efficiency will save

the cleaved strand and releases itself from the DNA.

you hours of time over other methods of cloning.
To harness the religating activity of topoisomerase, TOPO®

The technology behind TOPO® Cloning

vectors are provided linearized with topoisomerase I

The key to TOPO®Cloning is the enzyme, DNA topoiso-

covalently bound to each 3´ phosphate. This enables the

merase I, which functions both as a restriction enzyme and

vectors to readily ligate DNA sequences with compatible

as a ligase. Its biological role is to cleave and rejoin DNA

ends (Figures 1, 2, and 3) (1). In only 5 minutes at room

during replication. Vaccinia virus topoisomerase I specifi-temperature, the ligation is complete and ready for

cally recognizes the pentameric sequence 5´-(C/T)CCTT-3´

transformation into E. coli.

and forms a covalent bond with the phosphate group of the
Figure 1 - TOPO TA Cloning® of Taq-amplified DNA
Topoisomerase I recognition sites
5 minutes at
room temperature

AGGG
TTCCC

TOPO

P

P

3´ phosphate

CCCTT
GGGA

+

APCR Product

TOPO

A

CCCT T
GGGA A

TOPO

PCR Product

A AGGG

T TCCC

Topoisomerase I
is released

TOPO

TOPO TA Cloning® vector

Taq-amplified PCR product

Ligation complete

Figure 2 - Zero Blunt TOPO Cloning of blunt-end DNA
®

®

Topoisomerase I recognition sites
5 minutes at
room temperature

AAGGG
TTCCC

TOPO

P

P

CCCTT
GGGAA

3´phosphate

+

TOPO

PCR Product

CCCTT
GGGAA

TOPO

PCR Product

AAGGG
TTCCC

Topoisomerase I
is released

TOPO

Zero Blunt® TOPO® vector

Blunt-end PCR product

Ligation complete

Figure 3 – Directional TOPO Cloning of blunt-end DNA
®

Topoisomerase I recognition sites

P

5 minutes at
room temperature

AAGGG
TTCCC

TOPO

P

CCCTT
GGGAAGTGG

3´...