Answer to Question #118889 in Biochemistry for cieee

Question #118889
d. During the isolation of TOL plasmid from Pseudomonas putida, it was observed that the concentration of the plasmid DNA per 50 mL of sample was extremely low. Describe a technique you could employ to increase the concentration of the TOL plasmid.
Expert's answer

The concentration of the TOL plasmid can be increased using the polymerase chain reaction (PCR). In PCR, the target DNA is mixed with two primers (in excess), a thermostable DNA polymerase, and four deoxynucleotides. The PCR process involves three cycles; denaturation, renaturation, and synthesis of DNA.

First, the reaction mixture is placed in a high temperature of about 95 oC for one minute so the double-stranded DNA molecule can denature and form two single-stranded molecules. Then the temperature is reduced to allow for DNA annealing or renaturation. In this stage, the single DNA molecule will complementary pair with the primer. Lastly, initiation of DNA synthesis occurs at 3'-OH end of each primer.

Need a fast expert's response?

Submit order

and get a quick answer at the best price

for any assignment or question with DETAILED EXPLANATIONS!


Assignment Expert
30.05.20, 20:17

Dear Wendy, please post your question here to get an answer from the experts.

Wendy Dadson
29.05.20, 06:47

Why must the restriction enzyme used for the digestion of TOL plasmid be inactivated immediately after the restriction experiment? How could this be achieved?

Leave a comment

New on Blog