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Example sentences for: dttp

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Small samples (2 μg) from each RNA fraction were reverse transcribed and radiolabeled (2 μg oligo dT, 1x First Strand buffer, 5 mM DTT, 75 U SuperscriptII [Life Technologies], 20 mM dNTP mix containing dATP, dGTP, dTTP and 100 mCi 33P-dCTP (ICN Radiochemicals).
In the first DNA-protein binding reaction, [ 32P]-labeled double stranded oligonucleotides were generated by first annealing BSS2 (50 ng) to the BSS4 oligonucleotide (500 ng) then end-filling with 250 μM each of dATP, dGTP, and dTTP, 50 μCi of 32P-dCTP and Klenow.
The RT mix included 5.6 μl of 5X RT buffer, 0.02 mM of dNTPs (with dTTP, not dUTP, Perkin Elmer, Foster City, CA), 14.
To generate labeled single-stranded cDNA target, 10 μg total RNA from rat heart or brain (Clontech, Palo Alto, CA) was reverse transcribed for 2-3 h at 42°C in the presence of 6 μg random primers (Invitrogen/Life Technologies), 1x first-strand synthesis buffer (Invitrogen/Life Technologies), 10 mM DTT, dNTP mix (25 mM dATP, 25 mM dCTP, 25 mM dGTP, 15 mM dTTP, 10 mM amino allyl-dUTP), and 200 units Superscript II reverse transcriptase (Invitrogen/Life Technologies).
Reverse transcription was performed in the presence of 500 μM each of dATP, dCTP, and dGTP, 200 μM 5-aminoallyl-dUTP (Sigma), 300 μM dTTP, 1 × first-strand buffer, 10 mM dithiothreitol, and 400 U Superscript II (Life Technologies) in 40-μl reactions at 42°C for 3 h to overnight.