Ampicillin Resistance

A detailed Summary of Ampicillin Resistance

The purpose of this lab experiment was to see if a person could use a plasmid (R plasmid) vector containing certain fragments of foreign DNA for ampicillin resistance to be used to transform E. coli (Escherichia coli) cells to give the E. coli cells ampicillin resistance. Ampicillin resistance is needed in order for the E. coli to have the ability to survive in an ampicillin environment. Plasmid is a small ring of DNA that carries accessory genes separate from those of bacterial chromosome. Ampicillin is an antibiotic that is derived from penicillin that prevents bacterial growth by interfering with cell wall synthesis. E. coli is a commensal bacterium inhabiting the human colon that is widely used in biology, both as a simple model of cell biochemical function and as a host for molecular cloning experiments. In nature genes can be transferred between bacteria in different ways like transformation, conjugation, and transduction. One-way is bacterial transformation, it is when transfer of genetic information into a cell by the direct uptake of the DNA. Then the DNA is used to transform the cells from the original DNA to take a certain trait. How ever these bacteria can take up DNA only during the period at the end of logarit

5) Put both tubes on ice for 15 min(control). Then went the tubes are in the ice get two Luria agar plates and two Luria agar plates with ampicillin. Mark one of the plates + and the other - and then do the same for the plates with ampicillin.

Some of this data might not be completely accurate due to some errors. One error could be due to the fact that the temperature of the water bath could have been too high and killed or denatured the plasmids. One could also not of put the + and - tubes in the water bath or ice for the right amount of time. Also that there could have been contamination between the different substance that was used during the experiment or when spreading the E. coli on the different agar plates the bacteria spreader could of not been sterilized properly and contaminated the rest of the agar plates. As well when spreading the E. coli one could of broken up the agar in the agar plates and it could have changed the results. In addition one could have rushed and did not put the right amount of plasmids or Calcium Chloride into the solution, which might of change the results. There could be many things that could be the cause for the incorrect information.

6) Next take the tubes out of the waterbath and right away put them on ice for two minutes. (This part is critical to get right if not the plasmid would not of enter into he cells.)

In order for the plasmid to be able to enter the cell it must be heat shocked, so remove the tubes from ice and right away place them in to a 420C hot water bath for 60 to 90 seconds(control).

Some common words found in the essay are:
Ice Waterbath, Calcium Chloride, , DNA DNA, Results Discussion, Agar Platescontrol, agar plates, Luria Broth, bacteria growth, ampicillin plasmid, ampicillin resistance, bacteria spreader, plate ampicillin, growth agar, plates ampicillin, growth agar plates, agar plates ampicillin, incorporated coli, plasmid bacteria growth, plasmid bacteria, plate ampicillin plasmid, bacteria growth agar, Material Methods, Rubber Pipet, Graduated Pipets,

Approximate Word count = 1537
Approximate Pages = 6 (250 words per page double spaced)

Category: Science