A colleague in the lab wants to use a standard PCR to identify certain nucleic a

Question

A colleague in the lab wants to use a standard PCR to identify certain nucleic acid sequences from a retrovirus. You agree to the approach but suggest that RT-PCR be used instead. Why? The same colleague in question 3 is complaining that when carrying out a standard PCR on a patient's sample, she obtained an amplified DNA product even in the negative control tube (ie, no template DNA). You noticed that the samples were not handled properly and she wasn't wearing any gloves. What do you suspect is the problem?

Explanation / Answer

Solution to Question 3.

A retrovirus is a single stranded RNA virus.Standard PCR is used to amplify target DNA sequences.RT-PCR is used to amplify expressed genes by reverse transcription of the RNA in consideration of the retrovirus, into its DNA complement by the use of reverse transcriptase enzyme.Subsequently,the newly synthesized complementary DNA is amplified by the standard PCR technique.We thus suggest that RT-PCR be used.

Solution to question 4.

The standard PCR technique is a very sensitive technique.The negative control tube should not exhibit any amplified DNA product in normal cases.The presence of DNA product could be because of cross contamination.As it has been observed that the samples were not handled properly and she was not wearing any gloves,suggest a possible source of contamination in this case.

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