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

T4 RNA Ligase 2

T4 RNA Ligase 2

T4 RNA Ligase 2

T4 DNA Ligase

N/A
No ligase optimized for this activity
N/A
No ligase optimized for this activity
SplintR Ligase
100-1,000-fold higher efficiency than T4 DNA Ligase
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

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+
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/RNA
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.
T4 RNA Ligase 2
T4 RNA Ligase 2

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