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<br />Q.") <br /> <br />( <br /> <br />~.:.) <br />i..~ <br />l;,li <br />~ <br />~ <br /> <br />Endangered Fish Genetic Tissue Sampling Protocol <br />(San Juan 1991) <br /> <br />For allozyme analysis, ~ tissues will be taken from each Colorado <br />squawfish and razorback sucker: a muscle plug and caudal fin tissue. <br /> <br />( <br /> <br /><... <br /> <br />1.) All fish will be sampled non-lethally <br /> <br />2.) Anesthetize fish with appropriate concentration of MS-222 <br />3.) Cover fish with wet towel(s) to protect from wind and sun <br /> <br />4.) One person takes tissue, another person labels vials (dry hands <br />imperative) and records data <br /> <br />S.) Take tissue, <br /> <br />Use biopsy punches to cut and remove tissues. Scales do not have <br />to be removed prior to punching. One punch per tissue sample. <br />Tissue plug should be about 5 mm X 5 mm (or quarter inch by quarter <br />inch). If tissue is not removed with punch, it may have to be <br />snipped out with scissors and forceps. Keeping tissues clean, <br />place into 1.5 mm snap.cap vials. Swab tissue removal site with <br />Betadine. Clean and dry instruments between samples to avoid <br />contamination of tissues. If data collection complete, recover <br />fish and return to river. <br /> <br />6. ) <br /> <br />Label vials, <br /> <br />All important data cannot be placed on vial. coding is therefore <br />required. A three character number will be written on the vial. <br />The number will correspond to an individual fish. The number on <br />the vial will correspond to the PIT-tag number of the fish. The <br />PIT-tag number should then be recorded in the field notebook with <br />other pertinent data. On each vial, a short series of letters will <br />follow the number. The letter designations will correspond to <br />species and tissues: <br /> <br />Soecies <br /> <br />Tissue <br /> <br />RB razorback sucker <br />CS - Colorado squawfish <br /> <br />M - muscle <br />F fin <br /> <br />Example: The team collects its' first razorback sucker. Muscle is <br />placed in the first vial, which is labeled: OOlRBM. fin is placed <br />in the second vial, which is labeled: OOlRBF. Label vials with <br />permanent ink markers. The vial number and its associated tag <br />number (~) would be recorded in the field notebook. <br />7.) A standardized data sheet will be filled out completely at time of <br />capture. information will include: <br /> <br />fish number, PIT tag number <br />date collected <br />tissue collected <br />collectors name <br />capture location, river miles <br />fish length and weight <br />ecological data: depth, velocity, substrate, water <br />temperature <br /> <br />A4 <br />