Appendix I: Westslope cutthroat trout genetic sampling sites and results 



Wild Trout and Salmon Genetics Laboratory 



Division of Biological Sciences * University of Montana * Missoula, MT 59812 

 (406)243-5503/6749 Fax (406)243-4184 



December 2 1,2004 , 'T 



Ron Pierce . 



Genetics Contact, Region 2 



Mt.Dept. of Fish, Wildlife, and Parks ■ r ''p 



3201 Spurgin Road -« 



Missoula, MT 59801 ' -s 



Dear Ron: > 



'.-■"/-■.■■ 



The paired interspersed nuclear DNA elements (PINE) technique has been used to analyze DNA from the 

 following trout samples: . -. 



Table I. Summary of results. 



d e f 



Power (%) % WCT Individuals 



R94 Y85 100 xx 



'Number of samples successfully analyzed; if combined with previous sample (Indicated in "Location" column), number indicates the combined 



sample size; if present, the number in () is the average number successfully analyzed per locus (some individuals do not amplify for all marker 



loci). 



""Number of markers analyzed that are diagnostic for the non-native species. 



'Codes WSCT = westslope cutthroat trout (Oncorhyiiclws clarki lewisi); RBT = rainbow trout (O. mykiss); YSCT = Yellowstone cutthroat trout 



(O. clarki bouvieri). Only one species code is listed when the entire sample possessed alleles from that species only. However, it must be noted 



that we cannot definitively rule out the possibility that some or all of the individuals are hybrids; we merely have not detected any non-native 



alleles at the loci examined (see Power %). Species codes separated by "x" indicate hybridization between those species. 



■"Number corresponds to the percent chance we have to detect 1 % hybridization given the number of individuals successfully analyzed and the 



number of diagnostic markers used (e.g., 25 individuals are required to yield a 95% chance to defect 1% hybridization of rainbow or Yellowstone 



cutthroat trout into a westslope cutthroat trout population). Not reported when hybridization is detected. 



'Indicates the genetic contribution of westslope cutthroat trout to the sample assuming Hardy- Weinburg proportions. This number is reported 



only if samples appear to come from a random mating population and can be analyzed at the population level. 



"^Indicates number of individuals with genotypes corresponding to the species code column when the sample can be analyzed on the individual 



level only; this occurs when alleles are not randomly distributed and hybridization appears to be recent and/or if the sample appears to consist of 



an admixture of populations 



*See the "Sample Details" section below. " " " 



Methods and Data Analysis 



The PINE technique uses short synthetically made segments of DNA called primers, in pairs, to search for 

 relatively small segments of organismal DNA flanked by particular, often viral, DNA inserts. During the 

 polyTnerase chain reaction (PCR), the primers bind to the ends of the inserts and many copies of the organismal 

 DNA between the primers are made. While the DNA from some organisms may have two appropriately spaced 

 inserts to which the primers can attach, the DNA from other organisms may have only one or none of the 

 appropriately spaced inserts in particular regions. During PCR we will fail to copy DNA in the latter two cases. 



