Question

Paulo and Paula are two very enthusiastic foodies, writing the successful blog “Paulos for Paellas”.

In February 2020 the glamorous couple returned from Italy to Australia, bringing a selection of foods including cured meats and interesting cheese. Unfortunately, as they landed they were asked to wait along with the other passengers and submit bodily material for a genetic test for COVID-19. They were very angry to be held back for no apparent reason and have asked you, a friendly scientists taking their samples, to explain what is happening and what do you plan to do with their bodily samples.

3)Prepare a plan for RT-PCR analysis for COVID-19. Consider which controls to use and the plate layout.

Answer 1

Fo performing an RT-PCR let's use ubiquitin as the control in a plate

1st sample will be the negative control that is already tested negative for the corona virus.

2nd sample is your test sample

3rd sample is the sample of the already positive patient of the corona virus.

For each sample, we will put two reaction one using ubiquitin primer and the other one using corona virus-specific primers.

Now, from the samples, we have to isolate RNA and synthesize cDNA from this RNA.

Now, as the negative control, the coronavirus specific sample will not give any reading and its Cq value will be lower than the positive control. while the Cq value for actin will be the same for all the samples.

Now if you are infected then, corona virus specific primers will give Cq value almost equal to th positive or even more than that but if you are negative then your Cq value will be equally to the negative uninfected control sample.