'H-tm 'VO "-- ■ 



Sample 4 Water Name Location/Collection Date.' 

 Collector 



^ markers Species ID 



Power (•'.) %%VCT Indi'iduali 



3095 



3096 



3097 



3098 



3099 



Canyon Lake 



7 I4':0<)5 

 Ladd Kjiotek 



Patrick Creek 



- 29 :005 

 Laiid Kjiotek 



Webb Lake 



7 2l)2005 

 Ron Pierce 



Parker Lake 



7 21 2005 

 Ron Pierce 



West Twin Lakes 



6 21. 2005 

 Ron Pierce 



17 R6Y4 WCT? 



25 R6Y4 WCT.' 



R87Y74 100 a 



100 xx 



R6y4 WCT X RBT 



26 R6Y4 VCT X WCT X RBT 



2S R6Y4 WCT X YCT X RBT 



n 



dumber of fish successfully analyzed. If combined with a previous sample (Indicated in "Location" column), the number indicates the 



combined sample size. If present, the number in {) is the average number of individuals successfully analyzed per locus (some iiidividuals do not 



amplify for all marker loci). 



''Number of markers analyzed that are diagnostic for the non-native species (R=ra.inbow trout W=westslope cutthroat trout, Y= Yellowstone 



cutthroat trout). 



■^Codes: WCT = westslope cutthroat trout (Oncorhynchus clarki lewisi); RBT = rainbow trout (O. mykiss); YCT = 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 may not have detected any non-native alleles at 



the loci examined because of sampling error (see Power %). Species codes separated by "\" 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. For example, 25 individuals are required to yield a 95°'o chance to detect as little as 1% hybridization with 



rainbow or an 87% chance to detect as little as 1% hybridization with Yellowstone cutthroat trout into what once was a westslope cutthroat trout 



population. Not reported when hybridization is detected. 



Indicates the genetic contribution of the hybridizing taxa in the order listed under c to the sample assuming Hardy-Weinburg proportions. This 



number is reported if the sample appears to have come from a hybrid swarm. That is, a random mating population in which species markers are 



randomly distributed among individuals. 



Indicates number of individuals with genetic characteristics corresponding to the species code column when the sample can be analyzed on the 



individual level. This occurs when marker alleles are not randomly distributed among individuals and hybridization appears to be recent and'or if 



the sample appears to consist of a mixture of populations. 



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 

 polymerase 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 DN.A 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. 

 Thus, the PfNE technique coupled with PCR is used to search for evidence of genetic variation based on the 

 presence or absence of particular DNA fragments. The fragments are labeled by the primers used to produce 

 them and their length in terms of the number of nucleotides in the fragment. 



The fragments are made using dye labeled nucleotides and after PCR are separated from each other via 

 electrophoresis in polyacrylamide gels. Smaller fragments move through the gels at a faster rate than larger 

 fragments. The use of dye labeled nucleotides allows one to visualize the position of the fragments in the gels 



