DNA Assembly & Cloning Kits

ExSembly™ Cloning Master Mix

ExSembly™ cloning technology enables rapid, single-step, high-efficiency homology-based insertion of any amplified DNA product into a CIRCULAR vector. Vector linearization and DNA fragment insertion occur simultaneously, eliminating time-consuming linear vector preparation steps. Our innovative master mix buffer system supports robust restriction enzyme allowing vector linearization while maintaining high exonuclease and polymerase activities to enable homology-based DNA assembly. Successful fragment insertion destroys the vector-linearizing restriction enzyme site(s). Continuous restriction enzyme activity reduces negative empty-vector clones resulting in high-efficiency (typically >95%) fragment insertion.  ExSembly™ cloning mix in assembly cloning kit is compatible with most common restriction enzymes.
 

• One step: combines circular vector linearization and homology-based assembly.
• Rapid: saves 2-3 hours by eliminating vector digestion and gel purification steps.
• Efficient: Typically >95% of colonies bear the correctly inserted DNA fragment.
• Large scale: accommodates up to 500 ng plasmid DNA in one reaction, resulting in more colonies.
• Cost-effective: eliminates vector digestion, gel electrophoresis and purification costs, saving up to 50%.
 

Choose one (or more) restriction enzyme(s*) that linearize the vector and DO NOT cut within the fragment(s) to be inserted. For single fragment insertion, using PCR, add 20 bp homology upstream of the unique restriction site in the vector to the 5’ end of the fragment, and 20 bp of vector homology downstream of the restriction site to the 3' end.
 

Restriction enzyme compatibility

ORFs of eGFP or human SRSF3 were PCR-amplified and assembled into vectors (pQE-80L, pProex-hta, pRSET-B) using ExSembly™ Master Mix. ~200 ng vector DNA was used in each ExSembly™reaction. ExSembly™products were purified using a DNA mini-spin column and transformed to DH10B electrocompetent cells and plated on LB/Ampicillin plates. Forty colonies were picked from each reaction to perform colony PCR screening. Percent positive clones are shown in Figure 1 for each of 15 common restriction enzymes.

Product Name	Cat. #	Price	View
2× ExSembly™ Cloning Master Mix (10 rxns)	M0005	$169