Words similar to ddh
Example sentences for: ddh
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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.
The PCR products were purified with Qiagen PCR Purification Kit (Qiagen, Valencia, CA) and eluted into 42 μl of ddH 2 O, which was used for indirect labeling with PCR.
The two PCR products were diluted 1:500 in ddH 2 O and then hybridized to each other for 2 minutes at 95°C, 1 minute at 43°C and 1 minute at 72°C for 4 cycles.
A PCR reaction was set up with 1 μl of the reassociated DNA, 5 μl of 10x Clontech PCR buffer, 1 μl of 10 mM dNTPS, 2 μl of 10 μM CT primer or TT primer, 1 μl of Clontech cDNA polymerase and 40 μl of ddH 2 O. The reaction cycling conditions were 72°C for 5 min (for filling in the adapter ends), 94°C for 1 min, then 15 or 20 cycles of 94°C for 30 s, 66°C for 30 s, 72°C for 90 s, then 72°C for a final extension.
After phenol extraction and ethanol precipitation, the cDNA was resuspended in 25 μl of ddH 2 O. Both HL60 and HL60/TLK286 cDNA were digested with 3 μl (30 units) of the restriction enzyme Taq I in a final volume of 30 μl.