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Example sentences for: tlk

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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.
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.
Fifteen μl each of the adapterized HL60 cDNA and HL60/TLK286 cDNA were mixed with 30 μl of 2x HB buffer in a final volume of 60 μl, denatured at 95°C for 5 min and annealed at 68°C for 20 hours.
The denatured Cy3 (HL60) and Cy5 (HL60/TLK286) DNA were mixed and loaded onto a microarray chip.
For example, the stress kinase JNK1 was activated during TLK286 induced apoptosis [ 17 ] and JNK is known to be activated during apoptosis induced by a variety of stress stimuli [ 18 ] . However, in HL60/TLK286 resistant cells, we observed an overexpression of two phosphatases PP2 (AA599092) and MKP5 (AA056608) involved in JNK dephosphorylation and inactivation (Table 2).