nitrogen because of their tendency to thaw quickly. Frozen subsamples of blood and ground

heart and liver tissue were homogenized using QIAshredder spin columns (Qiagen Inc.). RNA

was then isolated with RNeasy Mini kits (Qiagen Inc.) using the manufacturer's protocol for

isolation of total RNA from animal tissues. DNA and RNA were isolated separately from a

minimum of three subsamples of each tissue from each turtle for a total of at least 18 subsamples

for each of 27 turtles unless tissue mass was insufficient.

 The concentrations of DNA and RNA in each subsample of blood, heart, and liver tissue

were determined using a PicoGreen dsDNA quantitation kit (Invitrogen Corporation, Carlsbad,

CA) and a RiboGreen RNA-specific quantitation kit with DNase I (Invitrogen Corporation) by

measuring fluorescence at standard fluorescein wavelength settings (485 nm excitation, 528 nm

emission) using a fluorescent microplate reader. Data were collected using KCJuniorTM data

analysis software.

 In addition, protein concentration of liver (but not heart or blood) was quantified for a

separate project (Chapter 4), so I included those data in the analysis of this study. Hepatic protein

concentrations were determined by Bradford assay (details, Chapter 4).

Statistical Analyses

 Data for body mass (BM) and carapace length (CL) were used to calculate specific growth

rates (SGR) for each turtle during the final 10-11 days of the study according to the following

equations:

 SGRbm = (ln[BMf] ln[BMi])*100/t
 SGRcl = (ln[CLf] ln[CLi])*100/t

where BMi and CLi represent body size 10 or 11 days prior to tissue sampling, BMf and CLf

represent body size on the day of tissue sampling, and t represents time (10 or 11 days).

Condition index (CI) was calculated as Fulton's K (CI = BMf/CLf3, Ricker 1975). Data for BMf,