Hello, I am having trouble doing a split plot analysis of variance with lme. The study design is ponds split in two, one side of each pond was a control and the other was one of three chemical treatments. Each pond side was sampled three times, response variable is the mean of twenty animals. So we have sample time within side within pond. Pond is random effects. There are three or four replicate ponds for each treatment data table looks like this pond side sampperiod response 1 cont 1 x1 1 cont 2 x2 1 cont 3 x3 1 chem1 1 x1 1 chem1 2 x2 1 chem1 3 x3 2 cont 1 x1 2 cont 2 x2 2 cont 3 x3 2 chem2 1 x1 2 chem2 2 x2 2 chem2 3 x3 3 cont 1 x1 3 cont 2 x2 3 cont 3 x3 3 chem3 1 x1 3 chem3 2 x2 3 chem3 3 x3 I can fit a model using aov m1<-aov(response~side*sampperiod + Error(pond/side/sampperiod, data=datafile) Since there is unequal replication, 4 replicates for one chemical treatment and 3 for the other two, and there is one missing value, lme from the nlme package should be used. m2<-lme(response~sampperiod*side*pond, random=~1|pond, data=datafile) Any help would be greatly appreciated. Cheers, Chris Edge PhD. Candidate Biology Dept. University of New Brunswick -- View this message in context: http://r.789695.n4.nabble.com/Help-with-lme-for-split-plot-tp3086237p3086237.html Sent from the R help mailing list archive at Nabble.com.