Method for clone checking.

Nader Nourizad; Baback Gharizadeh; Pål Nyrén

doi:10.1002/elps.200305434

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PubMed · 12783446

Method for clone checking.

Nader Nourizad·Baback Gharizadeh·Pål Nyrén

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Abstract

A new method for simple and fast clone checking is described. We combined the Pyrosequencing technology with a preprogrammed nucleotide dispensation strategy for fast analysis of DNA constructs. To test this method, the N-terminus region of plasmids constructed for the production of recombinant apyrase was analyzed. Of the ten plasmids tested, seven constructs were correct, two constructs showed one base deletion, and one construct showed deletion of a 195 bp fragment. The preprogrammed nucleotide dispensation strategy allowed the identification of the sequence downstream of the deletions. Thus, this method determines both the location and nature of possible artifacts.

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Apyrase Cloning, Molecular DNA DNA Mutational Analysis DNA, Recombinant Genetic Vectors Mutation Sequence Analysis, DNA Sequence Deletion Time Factors Journal Article Research Support, Non-U.S. Gov't

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Nader Nourizad, Baback Gharizadeh, Pål Nyrén. 2003. Method for clone checking.. https://doi.org/10.1002/elps.200305434

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