TAILED PRIMER FOR CLONED PRODUCTS USED IN LIBRARY CONSTRUCTION

摘要

This invention pertains to improved methods for the synthesis of long, double stranded nucleic acid sequences containing difficult to clone or variable regions.

主权项

1. A method of constructing a DNA library from a clonally purified synthetic gene with decreased contamination with the wild type background, said method comprising selectively amplifying recombinant sequences, the method comprising:
a) truncating a gene fragment by cleaving off sequences from both terminal ends; b) inserting the truncated gene into a cloning vector; c) amplifying the cloned gene out of the vector using
i. tailed primers that serve to add back the 5′ cleaved regions andii. bridging primers that serve to introduce new sequences within the gene fragment; and d) amplifying the resulting amplicon using primers that target the terminal regions.