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<br />( <br /> <br />( <br /> <br />L <br /> <br />C';l <br />'-~) <br />(-~ <br />(1..1" <br />e;;, <br />CJ1 <br /> <br />PROCEDURE FOR COLLECTING GAMETES <br /> <br />1. <br /> <br />Exude gametes into 15 ml Falcon sterile conical tubes with Screw <br />caps (Becton-Dickinson #4 -2095 -5; Fisher Scientific # 05 - 527 -90) by <br />applying gentle pressure to the flanks of the fish. Duplicate <br />tubes would be nice, if possible. Be certain not to contaminate <br />sample with water or dirt. <br /> <br />2. Tissues may be placed immediately on dry ice or stored on wet ice <br />for a few days (maximum 4-6) until they can be flash-frozen (using <br />dry ice). Note: these tubes become brittle when frozen at -80oC <br />and are more likely to crack. Be careful not to compress or <br />physically abuse tubes when they are this cold, as it could cause <br />them to shatter, spoiling the contents. <br /> <br />3. Store frozen tissues at -80oC (preferred - may use dry ice if <br />desired) or -20oC (for short periods) prior to shipment. Be <br />certain the tissues are not allowed to thaw. Thawing will result <br />in degradation of DNA and substantial reductions in yields. <br /> <br />4. Ship tissues on dry ice by overnight air. Be certain to include <br />enough dry ice to insure arrival without thawing (typically 5-10 <br />lbs. ) <br /> <br />PROCEDURE FOR COLLECTING TISSUES FROM DECEASED INDIVIDUALS <br /> <br />1. <br /> <br />Source tissues for mtDNA for dissected specimens are (in order of <br />best results): gonads,. heart, liver, spleen, kidney, muscle. <br />Dissect tissues (1-5 grams) and place in Falcon tubes (see above <br />for catalog information) or small screw cap tubes (as for allozyme <br />tissues). As above, duplicate tubes would be nice if the tissue is <br />available. <br /> <br />2. <br /> <br />Freeze dissected tissues immediately on dry ice or -20oC. Once <br />frozen they may be stored at -80oC (or on dry ice) or -20oC (for <br />short periods) . <br /> <br />3. <br /> <br />Ship tissues on dry ice by overnight air. Be certain to include <br />enough dry ice to insure arrival without thawing (typically 5-10 <br />lbs. ) <br /> <br />A6 <br />