7Kb plasmid

 7Kb plasmid Research

Abstract

After confirming that there was a 7000 base pair plasmid within the species Deinoccocus Aquaticus. The Start of this project is to isolate the plasmid and then transform and reintroduce it intintoe species deinococcus and then to other cells as well. Pending that complete transformation there will be further information. 

Introduction

The gel electrophoresis of pre-pcr is still not showing anything when run on a gel. This is still being researched. The Thermoscciece kit for plasmid extraction does seem going well and we are starting to get cconsistent nunumbers. There was also the use of the primers formulasatic 7kb and that was shown in figure 1. Now the use of a long amp primer will be used so that the plasmid will show in much more detail than  has been to this point. 

Methods

Theare someeme changes to our standard poprocedure, right now the use of lysozyme seems to be causing the ph balance to be off, and it is not used prproperly. So the beginning step of addinng lysozyme will be taken out. The centricentrifuginge 3ml of culture is still to happen. tthere wille no multibuffer at this titimeut that may change. Switched to the thermoscience kit. 

Results

Figure 1: PCR 1% gel with gelers.

Figure 2: 1% gel showing after PCR was done with different primers

Discussion

Through our process of looking for this 7kb plasmid there have been some inconsistaines still not understude. For one there was about how our plasmid is not adhering to the frit when it is going through the filtration process. this was slightly disproven when there we aliquotes taken form the from of the procedure. From this the plasmid has been proven to be in the extraction, the problem seems to bee thebect that the plasmid extractions are still not showing on any gel. While with the PCR process, the band is evidein, as in figure 1. In Figures shown that we now have the ability to find and isolate the plasmid very easily. The primers that wil have the entire plasmid replicated are in the process of being used. As thee is noting on this at present that will be the next part of this project.