TABLE 3.

Primers used in this work^a

Primer description	Name and 5′ sequence	Name and 3′ sequence
Outer	9A, CGCCTGAGTAGTACGTTCGC	3A, GCGGTGTGTACAAGACCCGA
Inner	8A, *TGGTGCATGGTTGTCGTCAG	5A, GAACGTATTCACCGCAGCATA
	7A, *GCATGGTTGTCGTCAGCTCG	5B, GAACGTATTCACCGTAGCGTA
		5F, GAACGTATTCACCGCGACATA
		5G, GAACGTATTCACCGCGACATG
Sequencing	SEQ1, Cy5-GACCCGAGAACGTATTCACC

^aThe asterisk shows that 8A and 7A were biotinylated at their 5′ end. The biotin interfered with the binding of primer 8A, but this problem was overcome using a hexaethylene glycol spacer. Primer 7A worked with or without the spacer. The sequencing primers were labeled with the fluorescent dye Cy5 and were designed to bind to the nonbiotinylated end of the PCR product. The sequences obtained were reversed and were complementary to the coding strand. The PCR amplification product from the outer primer pair was about 510 bp, and that from the inner pair was about 320 bp, depending on the organism.