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Example sentences for: dttp

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To generate the probe of HL60 DNA, a PCR reaction was set up with 0.5 μl of the reassociated DNA, 5 μl of 10x Clontech PCR buffer, 1 μl of dNTPS (10 mM dATP, dTTP, and dGTP each, 6 mM dCTP), 4 μl of FluoroLink Cy3-dCTP (Amersham Pharmacia), 4.5 μl of 10 μM CT primer, 1 μl of Clontech cDNA polymerase and 34 μl of ddH 2 O. In the PCR reaction for the probe of HL60/TLK286 DNA, Cy5-dCTP and TT primer were used instead of Cy3-dCTP and CT primer.
Subsequently, 25 μl of a premix containing 2 μl of 100 mM DTT (dithiothreitol), 10 μl 5X Superscript II buffer, 1 μl 50X aadNTP mix (25 mM dA, C, and GTP, 8.4 mM dTTP, and 16.
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).
Interestingly, the p66 INS/p51 INSmutant carrying insertion in both the subunits did not display the same reduction in dTTP binding affinity.
Reverse transcriptase activity in viral cores was assayed by incubating cores in endogenous polymerase buffer with 40 μM dATP, dCTP, dTTP and 2 μCi [α 32P]dGTP (Amersham) at 37°C for 90 min.