NEB DNA Polymerase
Thermophilic DNA Polymerases
| Vent | Vent (exo-) | Deep Vent |
Deep Vent (exo-) |
Taq | 9°Nm | Phusion** | LongAmp Taq |
LongAmp Hot Start Taq |
OneTaq | Phusion Hot Start Flex** | |
| Exonuclease Activities | |||||||||||
| 3'—>5' proofreading exonuclease |
yes | no | yes | no | no | no* | yes | yes | yes | yes | yes |
| 5'—>3' exonuclease activity |
no | no | no | no | yes | no | no | yes | yes | yes | no |
| Thermal Stability | |||||||||||
| Half life at 95°C | 6.7 hours | 6.7 hours | 23 hours | 23 hours | 0.9 hours | 6.7 hours | >6 hours at 96°C |
ND | ND | ND | ND |
| Half life at 100°C | 1.8 hours | 1.8 hours | 8 hours | 8 hours | <0.1 hours | 1.8 hours | 2 hours at 98°C |
ND | ND | ND | ND |
| Primer Extension Characteristics | |||||||||||
| Units/100 µl Primer Extension Rxn |
1-2 units | 2-4 units | 1-2 units | 2-4 units | 2-5 units | 1-2 units | 1-2 units | 10 units | 10 units | 2-5 units | 1-2 units |
| Error Rate (x10-6 base insertions) |
57 | 190 | ~20 | ~190 | 285 | in progress | 0.44 | ~140 | ~140 | ~140 | 0.44 |
| Resulting DNA Ends | blunt ends | 3´ A | blunt ends | 3´ A | 3´ A | 3´ A blunt |
blunt | 3´ A blunt |
3´ A blunt |
3´ A blunt |
blunt ends |
| Strand Displacement | yes; temp. dependent |
yes; temp. dependent |
yes; temp. dependent |
yes; temp. dependent |
no |
yes |
no | yes; temp. dependent |
yes; temp. dependent |
yes; temp. dependent |
no |
| Longest Primer Extension to Date |
13.2 kb | 15.0 kb | 14.0 kb | 15.0 kb | 8.1 kb | 14.0 kb | 37 kb | 40 kb | 40 kb | 15.0 kb | 20 kb |
- 1-5% of wild type exonuclease activity is present in the modified enzyme
Properties of DNA Polymerases
| 5'—>3' Exonuclease | 3'—>5' Exonuclease | Error Rate(x10-6) | Strand Displacement | Nick Translation | Thermal Stability | Km dNTPs | Km DNA | Extend RNA Primer | Extension from Nick | Primary Applications | |
| 9°Nm DNA Polymerase |
– | + | +++ | – | ++++ | 80 µM | 0.05 nM | – | + | Primer Extension | |
| Bst DNA Polymerase, Full Length |
+ | – | _ | + | + | + | + | ||||
| Bst DNA Polymerase, Large Fragment |
– | – | ++++ | – | + | + | + | Strand Displacement Applications | |||
| Bsu DNA Polymerase, Large fragment |
– | – | ++ | – | – | Labeling, 2nd Strand Synthesis, Strand Displacement | |||||
| Crimson Taq DNA Polymerase |
+ | – | _ | + | ++ | – | + | PCR (routine) | |||
| Phusion Hot Start Flex DNA Polymerase* |
– | ++++ | 0.44 | – | – | +++ | – | – | PCR (high fidelity, hot start) | ||
| Deep Vent DNA Polymerase |
– | +++ | ++ | – | ++++ | 50 µM | 0.01 nM | – | + | PCR (high fidelity) | |
| Deep Vent (exo–) DNA Polymerase |
– | – | +++ | – | ++++ | – | + | PCR (long) | |||
| E. coli DNA Polymerase I |
+ | ++ | 9 | _ | + | – | 1-2 µM | 5 nM | + | + | Nick Translation |
| Klenow Fragment 3´→ 5´ exo– |
– | – | 100 | +++ | – | – | + | + | Labeling | ||
| Klenow Fragment DNA Polymerase I |
– | ++ | 18 | ++ | – | – | 2 µM | + | + | Polishing Ends | |
| LongAmp Taq DNA Polymerase |
+ | ++ | ~140 | _ | + | ++ | – | + | PCR (routine, long) | ||
| LongAmp Hot Start Taq DNA Polymerase |
+ | ++ | ~140 | _ | + | ++ | – | + | PCR (hot start, long) | ||
| M-MuLV Reverse Transcriptase |
– | – | +++ | – | – | 18 µM | cDNA Synthesis | ||||
| OneTaq DNA Polymerase |
+ | ++ | _ | + | ++ | – | + | PCR (routine, difficult) | |||
| OneTaq Hot Start DNA Polymerase |
+ | ++ | _ | + | ++ | – | + | PCR (hot start, routine, difficult) | |||
| phi29 DNA Polymerase | – | ++++ | +++++ | – | – | 0.5 µM | + | + | Strand Displacement Applications | ||
| Phusion High-Fidelity DNA Polymerase* |
– | ++++ | 0.44 | – | – | +++ | – | + | PCR (high fidelity) | ||
| Sulfolobus DNA Polymerase IV |
– | – | – | – | + | DNA Synthesis Across Template Lesions | |||||
| T4 DNA Polymerase | – | ++++ | <1 | – | – | – | 2 µM | + | – | Polishing Ends, 2nd Strand Synthesis | |
| T7 DNA Polymerase | – | ++++ | 15 | – | – | – | 18 µM | 18 nM | + | – | Site Directed Mutagenesis |
| Taq DNA Polymerase | + | – | 285 | _ | + | ++ | 13 µM | 2 nM | – | + | PCR (routine) |
| Therminator DNA Polymerase |
– | – | + | – | ++++ | + | + | Chain Terminator Applications | |||
| Therminator II DNA Polymerase |
– | – | + | – | ++++ | + | + | Incorporation of Modified Nucleotide Analogs | |||
| Therminator III DNA Polymerase |
– | – | + | – | ++++ | + | + | Incorporation of Modfied Nucleotides | |||
| Therminator γ DNA Polymerase |
– | – | + | – | ++++ | + | + | Incorporation of gamma- phosphate Labeled deoxynucleotides | |||
| Vent DNA Polymerase |
– | ++ | 57 | ++ | – | +++ | 60 µM | 0.1 nM | – | + | PCR (routine, high fidelity) |
| Vent (exo–) DNA Polymerase |
– | – | 190 | +++ | – | +++ | 40 µM | 0.1 nM | – | + | PCR, Sequencing |
| 3'->5' Proofreading | Strand Displacement | Primary Applications | |
| Mesophilic DNA Polymerases | |||
| phi29 DNA Polymerase | ++++ | +++++ | Strand Displacement Applications |
| T4 DNA Polymerase | +++++ | - | Polishing Ends, 2nd Strand Synthesis |
| DNA Polymerase I | ++ | -* | Nick Translation |
| DNA Polymerase I, Klenow Fragment | ++ | ++ | Polishing Ends |
| Klenow Fragment (3' -> 5' exo-) | - | +++ | Labeling |
| T7 DNA Polymerase (unmodified) | ++++ | - | Site Directed Mutagenesis |
| Terminal Transferase | - | NA | 3' terminal Tailing |
| Thermophilic DNA Polymerases | |||
| Phusion® High Fidelity DNA Polymerase | +++++ | - | PCR (high fidelity) |
| Phusion® Hot Start High Fidelity DNA Polymerase | +++++ | - | Hot Start PCR (high fidelity) |
| PhusionHot® Start II High Fidelity DNA Polymerase | +++++ | - | Hot Start PCR (high fidelity) |
| Phire® Hot Start DNA Polymerase | + | - | Hot Start PCR (fast yield) |
| Phire® Hot Start II DNA Polymerase | + | - | Hot Start PCR (fast yield) |
| DyNAzyme™ EXT DNA Polymerase | + | + | PCR (difficult or long) |
| DyNAzyme™ II Hot Start DNA Polymerase | - | - | Hot Start PCR |
| Taq DNA Polymerase | - | -* | PCR (routine), Primer Extension |
| LongAmp Taq DNA Polymerase | + | -* | PCR (long, high-fidelity) |
| Crimson Taq DNA Polymerase | - | -* | PCR (routine) |
| VentR DNA Polymerase | +++ | ++ | PCR (high fidelity), Primer Extension |
| VentR (exo-) DNA Polymerase | - | +++ | PCR, Sequencing |
| Deep VentR DNA Polymerase | +++ | ++ | PCR (high fidelity), Primer Extension |
| Deep VentR (exo-) DNA Polymerase | - | +++ |
PCR (long), Primer Extension |
| 9°Nm DNA Polymerase | + | +++ | Primer Extension |
| Therminator™ DNA Polymerase | - | + | Chain Terminator Applications |
| Therminator™ II DNA Polymerase | - | + | Incorporation of modified substrates |
| Bst DNA Polymerase, Large Fragment | - | +++ | Strand Displacement Applications |
| Other Polymerases | |||
| M-MuLV Reverse Transcriptase | - | +++ | cDNA Synthesis |
| AMV Reverse Transcriptase | - | +++ | cDNA Synthesis |
| E. coli Poly(A) Polymerase | - | NA | 3´ labeling of RNA |
| phi6 RNA Polymerase | - | NA | dsRNA Synthesis |
| T7 RNA Polymerase | - | NA | High Yield RNA Production |
| SP6 Polymerase | - | NA | RNA Production |
- Degrades displaced strand.