Purified products were sequenced in separate reactions with each M13 primer using a cycle sequence of 96C, 10 sec; 50C, 5 sec; 60C, 4 min with BigDye® Terminator Mix v1.1 (Applied Biosystems, Foster City, CA).
The recovered DNA samples were subjected to sequencing with the matching T11N anchoring primer, using a cycle sequence technique in the presence of a radiolabeled terminator (USB Corp., Cleveland, OH, USA).
To analyse end sequences, BAC DNAs were amplified using a TempliPhi large construction kit (GE Healthcare, UK), and the end sequences were analysed according to the Sanger method using a cycle sequencing kit (Big Dye-terminator kit, Applied Biosystems, USA) with DNA sequencer type 3730xl (Applied Biosystems).
Sequencing was performed using a cycle sequencing kit (Big Dye-terminator Cycle Sequencing kit, Applied Biosystems) with DNA sequencer type 3730xl (Applied Biosystems) or DeNOVA-5000HT (Shimadzu Co., Japan) according to the protocols recommended by the manufacturers.
These PCR products were then sequenced using a cycle sequencing kit (Beckman Coulter, High Wycombe, UK) and run on an automated DNA sequencer (CEQ2000 Beckman Coulter).
DNA sequence analysis was performed using a cycle sequencing kit with dye-labeled terminators (PE Advanced Biosystems, Inc., Foster City, CA).
PCR samples exhibiting varying band shifting patterns were further purified using a High Pure PCR Product Purification kit (Boehringer Mannheim, Mannheim, Germany) and directly sequenced using a cycle sequencing protocol to determine precise variant sites.
Templates were sequenced by using a cycle sequencing reaction with fluorescent dye terminators (Perkin-Elmer, Applied Biosystems Division, Foster City, CA, USA), and the reaction products were analyzed by using an ABI 3100 (Perkin-Elmer) automatic sequencer.
PCR products were sequenced by using a cycle sequencing reaction with fluorescent dye terminators (Perkin-Elmer, Applied Biosystems Division, Foster City, CA, USA), and reaction products were analyzed with an ABI 3100 (Perkin-Elmer) automatic sequencer.
PCR products of positive samples were sequenced directly by using a dideoxynucleotide cycle sequencing method with an automated DNA sequencer (ABI PRISM 377; Perkin-Elmer).
After reamplification with the same set of primers the PCR products were sequenced on a semi-automated sequencer (ABI 3100, Applied Biosystems) using a Taq cycle sequencing kit (Applied Biosystems, Villebon Sur Yvette, France).
