DNA Ligase Selection Chart

While more than one ligase may work for your application, the following selection chart presents our recommendations for optimal performance. To select a ligase based upon the type of substrate present, please visit Substrate-based Ligase Selection Chart.  In addition, the Properties of DNA and RNA Ligases will help you understand the differences between them.


Blunt/TA
Ligase
Master Mix
Instant Sticky-end Master Mix ElectroLigase® T4 DNA Ligase Hi-T4™ DNA Ligase   Immobilized T4 DNA Ligase Salt-T4® DNA Ligase  Quick
Ligation™
Kit
NEBridge™ Ligase Master Mix  T3 DNA
Ligase
T7 DNA
Ligase
HiFi Taq DNA Ligase
Taq DNA
Ligase
9°N™ DNA
Ligase
NEBNext Quick Ligation™ Module SplintR® Ligase  E.coli DNA Ligase
DNA APPLICATIONS
Ligation of sticky ends ★★ ★★★ ★★ ★★  ★★ ★★★  ★★
★★★ ★★ ★★ ★ 
   ★
Ligation of blunt ends ★★★ ★★ ★★  ★★   ★★
★★★   ★★
 


   
T/A cloning ★★★ ★★ ★★  ★★   ★★
★★    


   
Electroporation

★★★ ★★  ★★  

 

 


   
Golden Gate Assembly       ★★★  a       ★★★                
Ligation of sticky ends only



     
 
★★★  


   
Repair of nicks in dsDNA ★★ ★★ ★★ ★★★  ★★★   ★★★
★★   ★★ ★★ ★★  ★★ ★★
★★ ★★ 
High-complexity library cloning ★★ ★★ ★★ ★★★       ★★  

 


   
Adapter Ligation  ▲
★★★ ★★ ★★       ★★  
 

   
Ligation-Dependent DNA Sequence & SNP Detection (LCR, LDR & related methods)



     
 

 ★★★ ★★ ★★
   
Ligation-Dependent RNA Sequence & SNP Detection


     
 

 


★★★  
Ligation of adjacent ssDNAs on an RNA splint



     
 

     
 ★★★  
NGS APPLICATIONS
NGS Library Prep dsDNA-dsDNA (ligation)

     
 
 

   
FEATURES
Salt tolerance
(>2X that for T4 DNA Ligase)




     
 
 


   
Ligation in 15 min. or less
       
 ✓  
Master Mix Formulation

     
 

 

   
Thermostable



     
 

 ✓
   
Thermotolerant



   
 

 ✓
   
Recombinant  ✓         ✓
Reusable/Removable



      
 






 
KEY
★★★ Optimal, recommended ligase for selected application
★★ Works well for selected application
Will perform selected application, but is not recommended
Please consult the specific NGS protocol to determine the optimal enzyme for your needs
 a Not yet tested 
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