FrontierScience / eeb31abb-41cb-4fe6-9864-f98fdcef338d / Molecular cloning is a technique that enables the generation of recombinant…

Problem

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Molecular cloning is a technique that enables the generation of recombinant plasmids that can induce exogenous expression of certain genes. The first step of molecular cloning is to reverse transcribe the mRNA of the gene of interest into single-stranded cDNA. In an experiment, this was achieved through the use of oligo dT primers that specifically anneal to the poly-A tail of mRNA molecules, which is then extended by reverse transcriptase enzyme. The resultant cDNA molecules are then amplified through PCR and the products were visualized using agarose gel electrophoresis to ensure that the gene of interest was correctly amplified. However, the observed DNA band was estimated to be ~1 kb in size, which is far below the expected 3.7 kb of the gene of interest. After some troubleshooting steps, it was determined that this discrepancy was due to the type of reverse transcriptase enzyme selected for the experiment. What specific characteristic of the enzyme can explain this discrepancy?

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biology

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Source and history

upstream_id / Original problem ID

eeb31abb-41cb-4fe6-9864-f98fdcef338d

task_id

22946681-50b5-5d9d-be71-25d36fbbad6b

task_key

olympiad--test--eeb31abb~2d41cb~2d4fe6~2d9864~2df98fdcef338d

task_revision_id

1

config / split

olympiad / test

Official source

initial import