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+
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 T3 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 T3 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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