The DNA fragment of the correct size was excised, purified with a Viogene gel extraction kit (VIOGENE, Sunnyvale, CA, USA), and sequenced directly by using a Prism automated DNA sequencing kit (Applied Biosystems, Foster City, CA, USA).
All constructs were confirmed by sequencing using a fully automated DNA sequencer (Beckman Coulter).
Sequencing products were purified by using Centri-Sep spin columns (Princeton Separations, Adelphia, NJ), and were resolved using an Applied Biosystems model 3130xl automated DNA sequencing system (Applied Biosystems) [9], [12], [13], [14].
Nucleic acid sequences of cDNA clones from the benign and malignant subtracted libraries were determined using an automated ABI 377 DNA sequencing system and standard dye terminator AmpliTaq kits (Perkin Elmer, Buckinghamshire, UK).
Sequence analysis was conducted using an ABI 377 automated DNA sequencer and Sequence Navigator software (ABI, Warrington, UK).
All SSH clones were selected and submitted to automated DNA sequencing using an ABI 3100 DNA analyzer, according to the manufacturer's instructions.
DNA sequencing was performed on the pretreated PCR product using an automated direct sequence analyzer (ABI PRISM 3100 Genetic Analyzer; Applied Biosystems, Foster City, CA, USA).
This was determined from the vector template using standard dye-labelled primer protocols from Applied Biosystems and a model 373A automated DNA sequencing system (Applied Biosystems, Inc., CA, USA).
Automated DNA sequencing was performed using a Beckman Coulter CEQ™ 8000 genetic analysis system (Beckman Coulter, Fullerton, CA) and analyzed using Beckman Coulter CEQ™ 8000 (Beckman Coulter) and Lasergene® software (DNASTAR, Madison, WI).
DNA sequencing was carried out using the BigDye Terminator v1.1 Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA) and separation of the products using an automated sequencing system (3130 Genetic Analyzer, Applied Biosystems).
