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Method for clone checking
Author(s) -
Nourizad Nader,
Gharizadeh Baback,
Nyrén Pål
Publication year - 2003
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305434
Subject(s) - clone (java method) , plasmid , computational biology , recombinant dna , fragment (logic) , apyrase , biology , nucleotide , pyrosequencing , genetics , nucleic acid sequence , sequence (biology) , dna , construct (python library) , computer science , microbiology and biotechnology , gene , algorithm , programming language , receptor
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.