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. The DNA Ligase Selection Chart helps you select the optimal DNA ligase for various applications. In addition, the Properties of DNA and RNA Ligases will help you understand the differences between them.

RECOMMENDED LIGASE COMMENTS
NICKED DNA/RNA
T4 RNA Ligase 2
T4 RNA Ligase 2
T4 RNA Ligase 2
T4 DNA Ligase
Immobilized T4 DNA Ligase
N/A No ligase optimized for this activity
T3 DNA Ligase  
SplintR Ligase 100-1,000-fold higher efficiency than T4 DNA Ligase
T4 DNA Ligase
Immobilized T4 DNA Ligase
For high temperatures, we recommend Taq DNA Ligase. For highest fidelity, we recommend HiFi Taq DNA Ligase.
ssDNA/RNA
N/A See CircLigase™
N/A No ligase optimized for this activity
T4 RNA Ligase 1 Supplement with ATP
T4 RNA Ligase 1 Supplement with ATP
T4 RNA Ligase 2 Truncated KQ
T4 RNA Ligase 2 Truncated KQ
Thermostable 5´ App DNA/RNA Ligase We recommend a Proteinase K cleanup
Thermostable 5´ App DNA/RNA Ligase We recommend a Proteinase K cleanup
RtcB Ligase Supplement with GTP and Mn2+
RtcB Ligase Supplement with GTP and Mn2+
T4 RNA Ligase 1
T4 RNA Ligase 1 Reported to work, but ligates inefficiently. Consider pdCp.
T4 RNA Ligase 1
T4 RNA Ligase 1
dsDNA
Blunt T/A Ligase Master Mix
Blunt T/A Ligase Master Mix
Quick Ligation Kit or Instant Sticky-end Ligase Master Mix For ligating ends under high salt conditions, we recommend Salt-T4 DNA Ligase. For ligation at temperatures up to 50°C, we recommend Hi-T4 DNA Ligase. For ligation of cohesive ends ONLY, we recommend T7 DNA Ligase.
Quick Ligation Kit or Instant Sticky-end Ligase Master Mix For ligating ends under high salt conditions, we recommend Salt-T4 DNA Ligase. For ligation at temperatures up to 50°C, we recommend Hi-T4 DNA Ligase. For ligation of cohesive ends ONLY, we recommend T7 DNA Ligase.

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