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Example sentences for: dttp

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Up to 50 ng of CIP-treated template DNA were incubated for 20 minutes at 37°C in a 10 μL solution containing 20 U TdT (NEB), 0.2 M potassium cacodylate, 25 mM Tris-HCl pH 6.6, 0.25 mg/ml BSA, 0.75 mM CoCl 2 , 4.6 μM dTTP and 0.4 μM ddCTP.
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 following reagents were added: 6 μl of 5X first strand buffer, 3 μl 0.1 M DTT, 0.7 μl 50X dNTP (25 mM dATP, dCTP, dGTP and 10 mM dTTP), 3 μl 1 mM Cy3-dUTP or Cy5-dUTP and 1.7 μl Superscript™ II (200 U/μl).
Reverse transcription was started by adding dNTPs to a final concentration of 1 mM dATP and 500 μM each: dCTP, dGTP, dTTP and 5-(3-aminoallyl)-2'-deoxyuridine-5'-triophosphate, (aa-dUTP) (Sigma), with 150 units of StrataScript (Stratagene, La Jolla, CA).
Radioactive products were made by 30 cycles of PCR amplification of 10-μl reaction volumes containing 10-30 ng of genomic DNA, 40 ng of each primer, 200 μM each of dATP, dTTP, dGTP, and dCTP, 1 μCi α- 32P-dCTP (3,000 Ci mmol -1) and 0.5 U Taq polymerase (Perkin Elmer).