. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 372 K. INDUE ET AL. 123 456. Figure 2. Representative results of amplification of the repetitive region of the adhesive protein gene. Amplified products were electro- phoresed on l% agarose gel. Lanes I and 2. Mylili\ eilitlis: lanes 3 and 4, M. irossuhts: lanes 5 and 6, M. galloprovincialis. M, molecular marker (lambda DNA digested with £«>T 141). inserted into the Sma I site of pUC19. Sequences of both strands of three independent clones were determined using a 373A DNA sequencer (Applied Biosystems Inc.) and a PRISM Dy
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 372 K. INDUE ET AL. 123 456. Figure 2. Representative results of amplification of the repetitive region of the adhesive protein gene. Amplified products were electro- phoresed on l% agarose gel. Lanes I and 2. Mylili\ eilitlis: lanes 3 and 4, M. irossuhts: lanes 5 and 6, M. galloprovincialis. M, molecular marker (lambda DNA digested with £«>T 141). inserted into the Sma I site of pUC19. Sequences of both strands of three independent clones were determined using a 373A DNA sequencer (Applied Biosystems Inc.) and a PRISM Dyeterminator Cycle Sequencing Kit (Applied Biosystems Inc.). Results Variation in the repetitive region The primers Me 13 and Me 14 were designed to amplify the repetitive region using the sequences identical to both M. edulis and M. galloprovincialis. Since the sense primer. Me 13. corresponds to a part of the nonrepetitive region and the antisense primer. Me 14, to a part of the 3' un- translated region, the whole repetitive region is amplified by PCR. AI. edulis. M. galloprovincialis, and M. trossulus were collected at Delaware, Kamaishi, and Juneau, re- spectively. These sampling points are known to be "pure sites" at which no other species of the M. edulis complex is found (McDonald el at., 1991). We analyzed 8, 16, and 8 individuals of AI. edulis, M galloprovincialis, and M. trossulus, respectively, using primers Me 13 and Me 14. Since the repetitive region is relatively long and highly repetitive, it was difficult to amplify the whole repetitive region if the template DNA was insufficiently pure and long, but prominent bands were successfully obtained by using well-purified, high molecular weight DNA. Typical results are shown in Figure 2. Sizes of the band ranged from to kb. The fragments obtained from M. edulis were generally larger than those of the other two species. The sizes of bands in M. trossulus and M. galloprovincialis were similar but, on a
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