PCR with degenerate primers containing deoxyinosine fails with Pfu DNA polymerase.

PCR with deoxyinosine-containing primers using DNA polymerases with proofreading activity.

1Department of Dermatology, Wayne State University, Detroit, Michigan 48201; 2Department of Dermatology, Niigata University, Niigata, Japan As the application of PCR has expanded, more amplified products have been used for cloning and mutagenesis. Although Taq polymerase (from eubacterium Thermus aquaticus) is the most convenient and intensively studied enzyme to date, one of its drawbacks is a...

Cautionary note on the use of dUMP-containing PCR primers with Pfu and VentR DNA polymerases.

PCR amplification of an Escherichia coli gene using mixed primers containing deoxyinosine at ambiguous positions in degenerate amino acid codons.

Skoog.M.T. 3035 Suzuki.Y. 2935 Walsh,K. 3027 Sokolova,N.I. 3067 Takaku,H2935 Webb.C.E. 3159 Somma.M.P. 2817 Takeda.N. 2841 Whittier,G. 2979 Souhami.R.L. 2929 Tibbetts,C. 3089 Wieben,E.D. 2869 Sparkes,R.S. 2869 Timberlake.W.E. 3105 Wierzchowski,K.L. 2947 Spencer,C. 3152 TittgenJ. 2993 Willcocks.T.C. 3160 Spohr.G. 3081 Tolun.A. 3159 Wilson,W.D. 2979 Stanford,D.R. 2869 Trowsdale.J. 2921 Yasueda,H....

Selecting Degenerate Multiplex PCR Primers

Single Nucleotide Polymorphism (SNP) Genotyping is an important molecular genetics technique in the early stages of producing results that will be useful in the medical field. One of the proposed methods for performing SNP Genotyping requires amplifying regions of DNA surrounding a large number of SNP loci. In order to automate a portion of this method and make the use of SNP Genotyping more wi...

PCR fidelity of pfu DNA polymerase and other thermostable DNA polymerases.

The replication fidelities of Pfu, Taq, Vent, Deep Vent and UlTma DNA polymerases were compared using a PCR-based forward mutation assay. Average error rates (mutation frequency/bp/duplication) increased as follows: Pfu (1.3 x 10(-6)) < Deep Vent (2.7 x 10(-6)) < Vent (2.8 x 10(-6)) < Taq (8.0 x 10(-6)) < < exo- Pfu and UlTma (approximately 5 x 10(-5)). Buffer optimization experiments indicated...