Presentation Type
Poster
Study the relationship of biomass and number of animals with the composition of amplified DNA
Abstract
The purpose of this study is to determine whether the composition of amplified PCR product is correlated with animal biomass or number during DNA amplification. Three different samples: animal only (ants, annelids, nematodes, rotifers and tardigrades); mixture of animal and soil; mixture of animal, soil and ethanol were used. Terminal Restriction Fragment Length Polymorphism (T-RFLP) was applied to detect animal DNA. Primers 18S11M-18S2a and 18S11C-18S2a were used to compare the primer bias in PCR amplification. Annealing temperatures of 55°C and 50°C were introduced to detect the stringency of primers in PCR amplification. Results show that the percent of amplified PCR product is correlated with the number of animals, not the biomass. DNA samples of each animal were mixed in pairs to detect if any particular animal DNA was inhibited in PCR by others or not. In the combination of nematode and ant, nematode was both amplified at 55°C and 50°C using 18S11M-18S2a but not by 18S11C-18S2a. In the combination of nematode and annelid, nematode was amplified at 50°C but not at 55°C. The results suggest that both primers and temperatures affect the composition of amplified PCR product and provide valuable information for future animal composition studies.
Categories
Natural Sciences
Research Type
Thesis
Mentor Information
Dr. James Garey
Study the relationship of biomass and number of animals with the composition of amplified DNA
The purpose of this study is to determine whether the composition of amplified PCR product is correlated with animal biomass or number during DNA amplification. Three different samples: animal only (ants, annelids, nematodes, rotifers and tardigrades); mixture of animal and soil; mixture of animal, soil and ethanol were used. Terminal Restriction Fragment Length Polymorphism (T-RFLP) was applied to detect animal DNA. Primers 18S11M-18S2a and 18S11C-18S2a were used to compare the primer bias in PCR amplification. Annealing temperatures of 55°C and 50°C were introduced to detect the stringency of primers in PCR amplification. Results show that the percent of amplified PCR product is correlated with the number of animals, not the biomass. DNA samples of each animal were mixed in pairs to detect if any particular animal DNA was inhibited in PCR by others or not. In the combination of nematode and ant, nematode was both amplified at 55°C and 50°C using 18S11M-18S2a but not by 18S11C-18S2a. In the combination of nematode and annelid, nematode was amplified at 50°C but not at 55°C. The results suggest that both primers and temperatures affect the composition of amplified PCR product and provide valuable information for future animal composition studies.
