Optimizing Conditions for Cloning cDNA from Chicken Brain

Acoustic over stimulation can lead to hair cell loss in the auditory epithelium. Hair cell loss or damage in mammals often results in decreased auditory sensitivity which is irreversible. However for birds, hair cells which are damaged by acoustic trauma or ototoxic drugs may be replaced by regeneration. As the first step in determining the molecular mechanisms of hair cell regeneration, gene expression techniques including cDNA synthesis and cloning were attempted first with the chicken brain. The initial attempts at making cDNA libraries were elusive; therefore, experiments were performed to identify problems and enhance the efficiency of specific steps. These steps included RNA isolation, competent cell preparation, and cell transformation. The study results suggest RNA isolations can be increased with an augmented ethanol precipitation and include a shorter protocol for competent cell preparation which increases transformation efficiency. In addition, the study results imply a possible transformation advantage for thawing competent cells at lower temperatures.

Name thesis:degree_name

Master of Science

Discipline thesis:degree_discipline

Department of Biology

Year

1994

Author dc:creator

• Clark, Stephen

• Medical Sciences

Repository record dc:identifier

https://digitalcommons.wku.edu/theses/936

OAI identifier oai:identifier

oai:digitalcommons.wku.edu:theses-1939