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Example sentences for: dutp

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cDNA from experimental samples was generated by reverse transcription in the presence of Cy-5 dUTP, and was mixed with a reference pool of cDNA derived from 10 different cell lines (Brown laboratory control reference 'E'), labeled with Cy-3 dUTP.
The PCR mix per reaction included 10 μl SYBR Green buffer, 1.5 mM MgCl 2 , 1 μM dNTP mix (with dUTP), 0.5 μl CMV primer mix, 2.5 units Ampli Taq gold, 0.05 units of uracil-N-glycosylase (UNG, Epicentre, Madison, WI) that had been diluted 1:10 in UNG buffer, and molecular grade water to bring the volume up to 45 μl with the addition of 5 μl of cDNA or standard.
Amino-allyl dUTP was incorporated during the PCR step as described in [ 22 ] , and at http://www.microarrays.org.
For each hybridization, 2 μg of each purified sample of polyadenylated RNA was reverse-transcribed in the presence of fluorescently labeled nucleoside triphosphates (Cy3-dUTP for reference sample, Cy5-dUTP for experimental sample).
In situ detection of DNA fragmentation was performed by incorporation of fluorescein-12-dUTP at the 3'-OH ends of DNA using Terminal deoxynucleotidyl Transferase (TUNEL assay) (Apoptosis Detection System, Fluorescein, Promega, Madison, WI), as performed previously by this laboratory [ 7 26 ] . Nuclei were counterstained using propidium iodide (Sigma Chemical Co., St. Louis, MO).