April 18,2005 



Wild Trout and Salmon Genetics Laboratory 



Division of Bioloj^ical Sciences * University of Montana * Missoula, MT59812 

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



Ron Pierce 



Genetics Contact, Region 2 



Mt. Dept. of Fish, Wildlife, and Parks 



3201 Spurgin Road ' ^ 



Missoula. MT 59801 



Ron: 



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

 following trout samples: 



Summary of results. 



a b c d e r 



Sample # Water Namc/Location/Collcction Date/ N # markers Species ID Power (%) % WCT Inilividuals 



2977 



Collector 



Cabin Creek 



17N1IW13 

 7/13/2004 

 Ron Pierce 



26 



R6Y4 WCT X RBT.WCT 



XX 



"Number 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 individuals do not 



amplify for all marker loci). 



""Number of markers analyzed that are diagnostic for the non-native species (R=rainbow 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 wheii 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 "x" indicate hybridization between those sp)ecies. 



''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% chance to detect 1% hybridization with rainbow or 



an 87% chance to detect 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 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. 



