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A transcription assay is performed with a plasmid that contains a known promoter

ID: 54291 • Letter: A

Question

A transcription assay is performed with a plasmid that contains a known promoter that produces a 400 base transcript. You have 8 microgram/ml of a 3.5 kb plasmid to serve as template in a 25 microliter reaction volume (DNA base pairs have a mass of 660 daltons). To a mixture of transcription factors and the other three NTPs you add alpha-32P-CTP, which can be easily either as soluble radioactivity or TCA precipitable RNA transcripts. The transcripts accumulate linearly for about an hour and plateau at a total incorporation of 4.8 pMol of CTP.

                        How many templates were present in each reaction?

                        How many transcripts are produced in each reaction?

                        How many transcripts per template are produced?

Explanation / Answer

25 templates are present i.e. 1 template per mL, so 25 in 25 mL.

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