DNA fragments (with expected sizes of 300 bp for DDB1 and DDB2, 172 bp for DHFR, 348 bp for PCNA, 198 bp for cyclin E and 220 bp for β-actin) were purified with the Prep A gene DNA purification matrix kit (Biorad), and their sequences were determined according to the dideoxy chain-termination method [35] and were found identical to those published previously.
Sequencing reactions were carried out according to the dideoxy chain termination method [ 25], using the ABI Prism Kit PE Applied Biosystemss) for the automatic method with ABI PRISM 377 (PerkinElmer).
The inserts were then sequenced with the T7 forward and M13 reverse primers using a dideoxy sequence analysis on an ABI3130xl Genetic Analyzer apparatus (Applied Biosystems) with the BigDye Terminator v3.1 cycle sequencing kit (Applied Biosystems) according to the manufacturer's recommendations.
After enzymatic purification with exonuclease I and shrimp alkaline phosphatase, the HPV16 PCR products were sequenced by the fluorescent dye dideoxy termination method using an ABI Prism 377 DNA sequencer (PE Applied Biosystems, Foster City, CA, USA) according to the manufacturer's protocol.
These fragments were sequenced by the dideoxy chain-termination method using the Big Dye Terminator v3.1 cycle sequencing Kit (Perkin Elmer Applied Biosystems, Courtaboeuf, France) according to the manufacter's instructions.
Primers and nucleotides were eliminated prior to dideoxy sequencing by treatment of the PCR reaction with ExoI and FastAP (Fermentas), according to the manufacturer's instructions.
Three clones were sequenced 3 times in both orientations by the dideoxy method, using M13 universal primers and ET Dynamic Terminator for MegaBACE (GE Healthcare, Fairfield, CT, USA).
The nucleotide sequences of NA and HA DNAs were confirmed by the dideoxy method using ABI PRISM 377 DNA Sequencer (Applied Biosystems).
DNA sequencing was carried out by using the dideoxy method, and sequences were compared with previously published sequences from the global collection of serotype A strains (10 ).
Sanger and his coworkers used the dideoxy method to sequence human mitochondrial DNA.
The cDNAs were sequenced via the dideoxy sequencing method (Big Dye Terminator 3.0; ABI Prism, Applied BioSystem, Branchburg NJ) and analyzed on an ABI Prism 310 sequencer (Applied BioSystem).
