High sequence fidelity in a non-enzymatic DNA autoligation reaction

High sequence fidelity in a non-enzymatic DNA autoligation reaction.

The success of oligonucleotide ligation assays in probing specific sequences of DNA arises in large part from high enzymatic selectivity against base mismatches at the ligation junction. We describe here a study of the effect of mismatches on a new non-enzymatic, reagent-free method for ligation of oligonucleotides. In this approach, two oligonucleotides bound at adjacent sites on a complementa...

preliminary expression of high fidelity pfu dna polymerase encoding fragment

High-fidelity gene synthesis by retrieval of sequence verified DNA identified using high-throughput pyrosequencing

Supplementary Data High-fidelity gene synthesis by retrieval of sequence verified DNA identified using high-throughput pyrosequencing Mark Matzas*, Peer F. Stähler*, Nathalie Kefer, Nicole Siebelt, Valesca Boisguérin, Jack T. Leonard, Andreas Keller, Cord F. Stähler, Pamela Häberle, Baback Gharizadeh, Farbod Babrzadeh, George Church febit group, Im Neuenheimer Feld 519, 69121 Heidelberg, German...

Calcium-driven DNA synthesis by a high-fidelity DNA polymerase

Divalent metal ions, usually Mg2+, are required for both DNA synthesis and proofreading functions by DNA polymerases (DNA Pol). Although used as a non-reactive cofactor substitute for binding and crystallographic studies, Ca2+ supports DNA polymerization by only one DNA Pol, Dpo4. Here, we explore whether Ca2+-driven catalysis might apply to high-fidelity (HiFi) family B DNA Pols. The consequen...

DNA polymerase fidelity and the polymerase chain reaction.

High-fidelity DNA synthesis conditions are those that exploit the inherent ability of polymerases to discriminate against errors. This review has described several experimental approaches for controlling the fidelity of enzymatic DNA amplification. One of the most important parameters to consider is the choice of which polymerase to use in PCR. As demonstrated by the data in Tables 2 and 3, hig...