Abstract: Derivation of a quantifiable mechanism of generating stable cell lines of genetically altered HTE cells.

Purpose: The purpose of the experiment was to develop pLXSN 16 E6 mutant vectors and introduce them into HTE cells using a QIAGEN midiprep kit. Methods: We identified 6 mutants on E6, namely 118-122, L50G, 9-13, I1284, 8S9A10T and 146-151. Using agar plates infused with ampicillin, we plated the bacteria and harvested single colonies after a 24-hour incubation period. This starter culture allowed us to make a secondary culture using LB broth and ampicillin. After centrifuging the resultant culture we obtained pellets of the bacteria and used the Quiaen midiprep kit to isolate the required DNA from each of the six samples. At the end of the midiprep, we recorded the concentration of DNA in each sample with the help of a nanodrop. We are currently involved in the process of cultivating a packaging cell line in order to transfect the retro viral DNA into cells and introduce them into HTE cells after confluence is reached. We then change the HTE cell media until an absolute cell line is obtained. Results: I dont know exactly what to write for either- the only thing I can think of is to obtain a cell line that can be studied.

Conclusion: