What is the error rate of DNA polymerase?

What is the error rate of DNA polymerase?

Scientists have reported mutation rates as low as 1 mistake per 100 million (10-8) to 1 billion (10-9) nucleotides, mostly in bacteria, and as high as 1 mistake per 100 (10-2) to 1,000 (10-3) nucleotides, the latter in a group of error-prone polymerase genes in humans (Johnson et al., 2000).

Do polymerases have different error rates?

In addition to different error rates, polymerases generate different error profiles (Keohavong and Thilly 1989). For example, T4 and modified T7 polymerases show primarily transitions of G•C>A•T, while Taq polymerase preferential shows A•T>G•C (Keohavong and Thilly 1989).

What is the error rate of PCR?

After 30 cycles of PCR amplifying a 3 kb template, only 3.96 % of the product DNA molecules contain 1 (nucleotide) error each. This means that 96.04 % of the product molecules are entirely error-free.

What is the error rate of Taq polymerase?

For Taq polymerase, which has a total error rate of 1.8 × 10−4 errors/base/doubling, single-molecule sequencing was sufficient to detect all types of polymerase errors, including base substitutions, deletions and insertions.

What is the error rate for transcription?

Occasionally, this enzyme makes mistakes that lead to small changes in the instruction that is produced. These mistakes are rare, but because cells make thousands of mRNAs, a single human cell can make 10-100 transcription errors per second.

Why does DNA polymerase make mistakes?

DNA polymerase can make mistakes while adding nucleotides. It edits the DNA by proofreading every newly added base. Incorrect bases are removed and replaced by the correct base, and then a new base is added.

Does Taq polymerase have high fidelity?

One unit of Platinum® Taq DNA Polymerase, High Fidelity, is the amount of enzyme required to incorporate 10 nmoles of deoxyribonucleotide into DNA in 30 min at 74°C.

What is a thermostable polymerase?

A thermostable DNA polymerase is used in repeated cycles of primer annealing, DNA synthesis and dissociation of duplex DNA to serve as new templates. The theoretical amplification of template DNA, assuming reagents are not limiting and the enzyme maintains full activity, is 2n where n is the number of cycles.

What is a high fidelity polymerase?

High-fidelity PCR enzymes are used for applications requiring high accuracy during DNA amplification such as cloning, sequencing or mutagenesis. Thermo Scientific Phusion high-fidelity DNA polymerases were created by fusing a dsDNA-binding domain to a Pyrococcus polymerase–like proofreading enzyme.

Is Taq polymerase High Fidelity?

Some of these enzymes have 3′-5′ proofreading exonuclease activity, absent in Taq-pol, which promotes the checking and removing mismatched nucleotides during the polymerization (Kunkel 1992; Kunkel and Bebenek 2000) and, therefore, are considered as high-fidelity DNA polymerases.