Words similar to first-strand

Example sentences for: first-strand

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For the hybridization experiments, 12 μg total RNA was used for first-strand cDNA synthesis as follows: RNA was mixed with a mixture of random hexamer (pdN 6 ) oligonucelotides and oligo-(dT 20 ) at final concentration 125 μg/μl for each oligonucleotide.
The purified 5 μg aRNA was labeled with Cy3 or Cy5 by reverse transcription in a solution containing 10 μg of random hexamer, 4 μl first-strand reaction buffer, 2 μl 10 mM dithiothreitol (DTT; Gibco-BRL), 1 μl of 2 mM dATP, dGTP, dTTP and 1 mM dCTP, 1 μl SUPERase-in, (Amtion) and 200 u Superscript II reverse transcriptase (Gibco-BRL).
We indirectly labeled the hybridization probes by incorporation of amino-allyl modified nucleotides in a first-strand cDNA RT reaction.
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.
First-strand cDNA synthesis was primed with 4 μg of random nonamers (New England Biolabs) or 6 μg random hexamers (Life Technologies) by heating at 70°C for 10 minutes, snap-cooling in dry ice/ethanol for 30 sec, and incubating at room temperature for an additional 5-10 min.