Choosing a Cloning Method | Thermo Fisher Scientific

A number of different methods, technologies, and protocols are available for performing the actual cloning reaction. To choose the one that best meets your needs, you will need to analyze the current parameters of your experiment and plan ahead for any downstream procedures. This section discusses several highly efficient cloning technologies:

Restriction Enzyme Digestion & Ligation Cloning: is an industry standard for molecular biologists
TOPO Cloning: provides simple, convenient reactions typically in less than 5 min
Gateway Cloning Technology offers the greatest flexibility in vector choices and downstream applications
Seamless Cloning and Assembly: Tools optimized to clone up to 4 DNA fragments simultaneously into virtually any linearized E. coli vector in a 30 minute room temperature reaction
Type IIs Assembly: Clone up to 8 fragments simultaneously and without scars, and avoid rearrangements when cloning homologous or repetitive sequences
GeneArt Gene Synthesis and Strings DNA Fragments: 100% accuracy and optimization of genes to maximize expression

The table below can help you navigate your way through these technologies.

View our Cloning Technologies Guide

	Restriction Enzyme Cloning	TA-Cloning	TOPO (TA, blunt, directional)	Gateway Cloning Technology	GeneArt Seamless Cloning	GeneArt Type IIs Assembly	GeneArt High-Order Genetic Assembly
Fragments Cloned Simultaneously	1	1	1	Up to 4	Up to 4	Up to 8	Up to 10
Max Fragment(s) Size	Variable	1-3 kb	<5 kb (<10 kb for XL-TOPO)	Variable	Up to 10 kb, max total size of 40 kb	Up to 10 kb, max total size of 13-20 kb	Up to 100 kb, max total size of 110 kb
Gene Shuttling Between Vectors w/o PCR or Restriction Enzymes	NO	NO	NO	YES	NO	NO	NO
Seamless (No Extra Sequences)	NO	NO	NO	NO	YES	YES	YES
Use Your Own Vector	YES	NO	NO	YES (may require conversion)	YES	YES	YES
Time to Clone Multiple Fragments	Days to Weeks	Not Possible	Not Possible	>4 Days	1 hr	1 hr	3 Days
4 Fragment Cloning Efficiency	NA	NA	NA	30% - 85%	75%	>90%	>90%
Web-Based Vector Design Tool	NO	NO	NO	NO	YES	YES	YES

Browse for vectors using our Vector selection tool