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Enzymes for Molecular Biology

Labeling Nucleic Acids

There are two ways to label DNA with enzymes: at the ends or internally along the molecule. Detectable markers such as fluorophores or modified nucleotides can be introduced by the addition of 5’ and 3’ groups through enzyme action, as well as by introducing internal markers through direct incorporation or post-labeling techniques with a suitable enzyme.

Labeling by the action of an enzyme is required for many studies aiming to elucidate the functions and dynamics of nucleic acids in vitro and in cells. For every site where labeled nucleotides can be added to DNA or RNA, there is a suitable enzyme that can catalyze the addition.

Enzyme Activity Labeling action
    End
labeling
Direct
incorporation
By random
priming
Gap label Nick 
translation
P7060L
Klenow Fragment

DNA polymerase with 5’→3’
synthesis and 3’→ 5’
exonuclease activities

✘ No ✔ Yes ✔ Yes ✘ No ✘ No
P7010
Klenow (3’→5’ exo-)

DNA polymerase with 5’→3’
synthesis activities
 ✔ Yes (3’ dsDNA) ✔ Yes ✔ Yes ✘ No ✘ No
P7260L
T7 DNA polymerase
Coming soon
DNA polymerase with 5’→3’
synthesis and 3’→5’ ssDNA
and dsDNA exonuclease activity.
[The double-stranded DNA
exonuclease activity requires
the presenceof thioredoxin.]
 ✔ Yes (5’ dsDNA) ✔ Yes ✘ No ✔ Yes ✘ No
P7050L
DNA polymerase I
DNA polymerase with 5’→3’
synthesis, 5’→3’ and 3’→5’
exonuclease activities
✘ No ✘ No ✘ No ✘ No ✔ Yes
RP810
TaqNova Stoffel DNA
Polymerase
BLIRT
Stoffel Fragment derivatives of Taq
DNA polymerase that is 5’→3’
exonuclease deficient
✘ No ✔ Yes ✘ No ✘ No ✘ No
P7620L
TaqIT DNA
polymerase

TaqIT is 5’→3’ exonuclease
deficient derivative of Taq
DNA polymerase (Stoffel)
✘ No ✔ Yes ✘ No ✘ No ✘ No
P7070L
Terminal
deoxynucleotidyl
transferase (TdT)
Coming soon
Catalyzes the addition of dNTPs
to the 3' hydroxyl terminus of
DNA molecules; extends blunt,
5’-overhanging, ssDNA and RNA
 ✔ Yes (3’ ssDNA) ✘ No ✘ No ✘ No ✘ No
P7180L
T7 RNA polymerase
Coming soon
Synthesizes RNA in 5’→3’
direction off DNA template
✘ No  ✔ Yes (RNA) ✘ No ✘ No ✘ No
P7460L
Poly(A) polymerase

Catalyzes the addition of AMP
from ATP to the 3’ hydroxyl
of RNA for poly(A) tailed RNA
 ✔ Yes (3’ RNA) ✘ No ✘ No ✘ No ✘ No
L6050L
T4 RNA ligase 1
Coming soon
Ligates RNA and ssDNA
molecules
3’ (RNA and ssDNA) ✘ No ✘ No ✘ No ✘ No
Y9040L
T4 polynucleotide
kinase (PNK)

Catalyzes the transfer of the
γ-phosphate from ATP to
the 5´-terminus of
polynucleotides or to
mononucleotides bearing
a 5´-hydroxyl group
5’ (ssDNA, dsDNA
and RNA)
✘ No ✘ No ✘ No ✘ No
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