What is the principle behind agarose gel electrophoresis

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Reference no: EM133758419

PCR and DNA Gel Electrophoresis

Application of PCR and DNA gel electrophoresis in diagnostics:
Chronic Hepatitis is a condition of the liver that is caused by many different viruses. Of these, Hepatitis B (HBV) and Hepatitis C (HCV) have been identified as the top causes of hepatitis. Hepatitis B, C and D are very similar in their modes of transmission and are spread by contact with infected blood and contaminated syringes. Simultaneous co-infections with B and C can occur in patients. A group of patients (1-5) was tested for Hepatitis C which has an RNA genome. Briefly blood samples were taken and RNA was extracted. RNA samples were subjected to amplification by RT-PCR. The products were resolved by agarose gel electrophoresis.

The picture of the gel is shown below in Figure 1. L is the DNA standard; - and + are the negative and positive controls respectively; 1-5 are the RT-PCR amplified samples from 5 patients. A positive amplification of HCV and HCB are indicated by arrows.

Question 1: What is the principle behind agarose gel electrophoresis?

Question 2: Which patients are infected with HCV, HCV and both HBV and HCV (co-infection) and no infection? How did you make this conclusion?
Question 3: Using points A and B as reference, predict the direction that the DNA molecules moved in the gel. Did they move from A to B or from B to A?

Question 4: Briefly explain the steps that were done in Figure 2 to amplify the viral nucleic acid (you need not explain the agarose gel electrophoresis part). How did this differ from the amplification in Figure 1?

Question 5: What is the purpose of negative and positive controls in this experiment?

Reference no: EM133758419

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