Substrate-based Ligase Selection Chart

          

This chart provides our recommendation for a choice of ligase to use in a reaction, based upon the type of substrate present. DNA, RNA and hybrid substrates are represented and require specific enzymes to achieve the highest-efficiency ligation.


RECOMMENDED LIGASE
COMMENTS
NICKED DNA/RNA
Substrate_ligaseChart_Nicked1
T4 RNA Ligase 2

Substrate_ligaseChart_Nicked2 T4 RNA Ligase 2

Substrate_ligaseChart_Nicked3 T4 RNA Ligase 2

Substrate_ligaseChart_Nicked4 T4 DNA Ligase

Substrate_ligaseChart_Nicked5 N/A
No ligase optimized for this activity
Substrate_ligaseChart_Nicked6 N/A
No ligase optimized for this activity
Substrate_ligaseChart_Nicked7 SplintR Ligase
100-1,000-fold higher efficiency than T4 DNA Ligase
Substrate_ligaseChart_Nicked8 T4 DNA Ligase
For high temperatures, we recommend Taq DNA Ligase.
For highest fidelity, we recommend HiFi Taq DNA Ligase.
ssDNA/RNA
Substrate_ligaseChart_SingleStranded1 N/A
See CircLigase™
Substrate_ligaseChart_SingleStranded2 N/A No ligase optimized for this activity
Substrate_ligaseChart_SingleStranded3 T4 RNA Ligase 1
Supplement with ATP
Substrate_ligaseChart_SingleStranded4 T4 RNA Ligase 1

Substrate_ligaseChart_SingleStranded5 T4 RNA Ligase 2 Truncated KQ

Substrate_ligaseChart_SingleStranded6 T4 RNA Ligase 2 Truncated KQ

Substrate_ligaseChart_SingleStranded7 Thermostable 5´ App DNA/RNA Ligase
We recommend a Proteinase K cleanup
Substrate_ligaseChart_SingleStranded8 Thermostable 5´ App DNA/RNA Ligase We recommend a Proteinase K cleanup
Substrate_ligaseChart_SingleStranded9 RtcB Ligase Supplement with GTP and Mn2+
Substrate_ligaseChart_SingleStranded10 RtcB Ligase
Supplement with GTP and Mn2+
Substrate_ligaseChart_SingleStranded11 T4 RNA Ligase 1

Substrate_ligaseChart_SingleStranded12 T4 RNA Ligase 1 Reported to work, but ligates inefficiently. Consider pdCp.
Substrate_ligaseChart_SingleStranded13 T4 RNA Ligase 1
Substrate_ligaseChart_SingleStranded14 T4 RNA Ligase 1

dsDNA/RNA
Substrate_ligaseChart_DoubleStranded1 Blunt T/A Ligase Master Mix

Substrate_ligaseChart_DoubleStranded2 Blunt T/A Ligase Master Mix
Substrate_ligaseChart_DoubleStranded3 Quick Ligation Kit or Instant Sticky-end Ligase Master Mix For ligating ends under high salt conditions, we recommend T3 DNA Ligase. For ligation of cohesive ends ONLY, we recommend T7 DNA Ligase.
Substrate_ligaseChart_DoubleStranded4 Quick Ligation Kit or Instant Sticky-end Ligase Master Mix For ligating ends under high salt conditions, we recommend T3 DNA Ligase. For ligation of cohesive ends ONLY, we recommend T7 DNA Ligase.
Substrate_ligaseChart_DoubleStranded5 T4 RNA Ligase 2
Substrate_ligaseChart_DoubleStranded6 T4 RNA Ligase 2

CIRCLIGASE™ is a trademark of EpiCentre Technologies Corp.