A Method to Direct Sequence Cosmid LAWRIST16 Clones

Abstract

We developed a new cycle condition method optimized for direct sequencing LAWRIST16 cosmid clones based on dye terminator and dye primer cycle sequencing chemistries. We report a direct comparison of the two most widely used sequencing polymerase enzymes (AmpliTaq FS and Thermosequenase). The sequencing data obtained is of high quality and we believe this method could be routinely used in large scale sequencing facilities.

A new candidate region for the positional cloning of the XLP gene

“…Plasmid and cosmid end sequencing was performed using Dye Terminator sequencing chemistry with Taq polymerase together with T3/T7/Sp6 primers. 12 Primer walking using Dye Terminator sequencing chemistry was used to complete the sequence of each subclone in addition to confirming sequence overlaps of adjacent subclones. All sequences thus generated were assembled using Applied Biosystems programs FACTURA version 1.2.0 and AUTOASSEMBLER version 1.3.0.…”