Pglo Transformation Lab Report
In this lab, we performed a genetic transformation through the process of gene transfer. Gene transfer involves the insertion of a gene into an organism. The gene to be inserted is usually contained in a plasmid, which is relatively small, circular non-chromosomal DNA molecule typically found in bacteria. Once the plasmid containing the gene is inserted into the organism, it is absorbed into the organism’s own genetic code. After this occurs, the newly introduced gene begins coding for proteins, giving the organism a new trait. This process of genetic transformation is often used in the field of biotechnology. For example, genes that code for proteins that protect against herbicide are often added to crops such as corn so that farmers can target
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Coli on a plate of LB agar. We then put 250 µn of CaCl2 transformation solution into two micro test tubes for the purpose of changing the bacterium’s cell wall to allow the pGLO plasmid to enter more easily. Second, we placed one colony of E. Coli from our original plate into each of the micro test tubes. Subsequently, we extracted a loopful of pGLO plasmid with a sterile loop and then placed it in one of the micro test tubes. We incubated the tubes on the ice for ten minutes while we prepared four new agar plates. One plate for +pGLO E. Coli on agar with ampicillin, one plate for +pGLO E. Coli on agar with ampicillin and arabinose, one plate for -pGLO E. Coli on agar with ampicillin, and one plate for -pGLO E. Coli on plain LB agar. After ten minutes had passed, we heat-shocked the bacteria by transferring the tubes into a hot water bath for 50 seconds and then rapidly transferring them back to the ice. After the tubes had remained in the ice for two minutes, we removed them from the ice and added 250 µm of LB nutrient broth to them. Next, we let the tubes rest at room temperature for ten minutes. After ten minutes had passed, we added the corresponding bacterial suspensions to the plates and then placed the plates in a 37ºC incubator for twenty-four
Coli on a plate of LB agar. We then put 250 µn of CaCl2 transformation solution into two micro test tubes for the purpose of changing the bacterium’s cell wall to allow the pGLO plasmid to enter more easily. Second, we placed one colony of E. Coli from our original plate into each of the micro test tubes. Subsequently, we extracted a loopful of pGLO plasmid with a sterile loop and then placed it in one of the micro test tubes. We incubated the tubes on the ice for ten minutes while we prepared four new agar plates. One plate for +pGLO E. Coli on agar with ampicillin, one plate for +pGLO E. Coli on agar with ampicillin and arabinose, one plate for -pGLO E. Coli on agar with ampicillin, and one plate for -pGLO E. Coli on plain LB agar. After ten minutes had passed, we heat-shocked the bacteria by transferring the tubes into a hot water bath for 50 seconds and then rapidly transferring them back to the ice. After the tubes had remained in the ice for two minutes, we removed them from the ice and added 250 µm of LB nutrient broth to them. Next, we let the tubes rest at room temperature for ten minutes. After ten minutes had passed, we added the corresponding bacterial suspensions to the plates and then placed the plates in a 37ºC incubator for twenty-four