Na Transformation Lab Report
NA transformation of E. coli:
The two plasmids were added to individual tubes containing E. coli and one with no plasmids. The three samples of E. coli were heated in a 42°C water bath for 90 seconds to heat shock the bacteria so that the plasmids would be taken up by the E. coli. These samples were then incubated at 30°C for half an hour and then plated on LB agar. Each tube was plated on an LB plate and a LB + ampicillin plate. Ampicillin is an antibiotic that is effective against E. coli, both pSA11 and pDH223 contain a gene for ampicillin resistance, so if the plasmids undergo transformation with the E. coli then growth will be seen on the LB+Amp plates. The results of this step showed that the plasmids were transformed as growth was seen on both the LB and LB+Amp plates. …show more content…
coli samples were centrifuged to make a pellet, which was resuspended using glucose, then NaOH was added to lyse the cells, acetic acid was added to precipitate the cellular components. The mixture was then centrifuged for 10 minutes and the supernatant was transferred to a new tube. Isopropanol was then added to precipitate the plasmid DNA and centrifuged into a pellet, dried, and then resuspended with distilled water. This plasmid DNA was then run on an agarose gel to view the stands of
The two plasmids were added to individual tubes containing E. coli and one with no plasmids. The three samples of E. coli were heated in a 42°C water bath for 90 seconds to heat shock the bacteria so that the plasmids would be taken up by the E. coli. These samples were then incubated at 30°C for half an hour and then plated on LB agar. Each tube was plated on an LB plate and a LB + ampicillin plate. Ampicillin is an antibiotic that is effective against E. coli, both pSA11 and pDH223 contain a gene for ampicillin resistance, so if the plasmids undergo transformation with the E. coli then growth will be seen on the LB+Amp plates. The results of this step showed that the plasmids were transformed as growth was seen on both the LB and LB+Amp plates. …show more content…
coli samples were centrifuged to make a pellet, which was resuspended using glucose, then NaOH was added to lyse the cells, acetic acid was added to precipitate the cellular components. The mixture was then centrifuged for 10 minutes and the supernatant was transferred to a new tube. Isopropanol was then added to precipitate the plasmid DNA and centrifuged into a pellet, dried, and then resuspended with distilled water. This plasmid DNA was then run on an agarose gel to view the stands of