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By P. Michael Conn

Neuroendocrine Peptide method

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The amount of total RNA in each sample was determined from the standard curve. 5 ng/μΐ of sample. We found that at higher concen­ trations the results obtained with the spectrofluorometer were compara­ ble to results obtained with the spectrophotometer. To quantitate the amount of RNA isolated from nuclei preparations, one must ensure that the fluorescence detected in the sample is due to RNA and not to any DNA that may be remaining in the sample. This is especially important because DNA binds EtBr more efficiently, such that a small amount of DNA contamination in the sample can be a real prob­ lem.

PROBE wm — im m ·» PRIMARY TRANSCRIPT [ exi IN T R O N A Γ ^ Ι IN T R O N B " ■ aaaaa EXO N I S i NUCLEASE POSSIBLE PROCESSING INTERMEDIATES l E X I |EX II I IN T R 0 N B | EXO N III |A A A A EX II £ --------------------S i NUCLEASE SPLICED OUT INTRONA OR IE l} INTRONA E X II I I AAA A EXO N I EX II ~ 2 SPLICED OUT . INTRON B S -ι NUCLEASE MATURE mRNA j EX I I EX II | EXO N III mature mRNA in the nucleus J a AA AA A S-, NUCLEASE Γε χ ν Ί a c r y l a m id e g el PROBE c PRIMARY TRANSCRIPT < EXON II /INTRON B < mRNA (EX II) < INTRON A F ig .

The transfer of genes into the germ line of mammals such as mice to create transgenic animals is a very powerful tool for genetic studies and is de­ scribed elsewhere in this series. 50 N. Sarver, S. -F. Law, W. T. McAllister, J. C. Byrne, and P. M. Howley, in “Genetic Engineering” (J. K. Setlow and A. ), p. 173. Plenum, New York, 1983. [2] NEUROPEPTIDE GENES 35 Acknowledgments We would like to thank Dr. David Moore from the Department of Molecular Biology at Massachusetts General Hospital for providing the plasmid pOCAT.

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