Kinase Linkers

1. 0.8 pg of 32P-labeled linkers is needed for each cDNA preparation (see Note 4). An example reaction is the following:

2 pL 10 kinase buffer; 1 pL a-32P-ATP;

1 pL T4 polynucleotide kinase (~5 U); 8 pL H2O. Incubate for 15 min at 37°C.

1 pL 10X kinase buffer; 4 pL 10 mM ATP;

1 pL T4 polynucleotide kinase;

4 pL ddH2O. Incubate for 45 min at 37°C.

3. Heat-kill the enzyme at 80°C for 15 min, and then cool on ice.

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