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Worksheet IX.18
Joining DNA
molecules - Ligation
Table 1. Properties of ligases used in genetic engineering (Sambrook and Russel 2001, changed)
Substrates
Ligase
Optimal reaction
temperature
cohesive
ends
blunt
ends
DNA-RNA
hybrids
RNA-RNA
hybrids
Cofactors
E. coli DNA
ligase
Yes
Yes
No
No
NAD +
10-15 °C
(cohesive ends)
Bacteriophage
T4 DNA ligase
Yes
Yes
Yes
Yes
ATP
4 °C (cohesive
ends) 15-25 °C
(blunt ends)
DNA ligases of
thermophile
bacteria
Yes
No
No
No
NAD +
24-37 °C
(cohesive ends)
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