Random-primed labeling of DNA with Klenow


15.5 µl            DNA
4 µl                 oligo-dNTP mix
1 µl                  Tris/MgCl2 mix
1 µl                 100X BSA from NEB
1 µl                 Klenow (exonuclease deficient)
2.5 µl              alpha-32P dATP (Perkin Elmer, 6000 Ci/mmole, 10 mCi/ml)

Oligo-dNTP mix
1.25 M            HEPES, pH 6.6;
62.5 mM         b-mercaptoethanol
895 µg/ml     random hexamers (1 A260 unit = 26.5 µg)
125 µM each dCTP, dGTP, and dTTP (made from 100 mM stock solutions from Amersham-Pharmacia)

Tris/MgCl2 mix
625 mM Tris-HCl pH 8.0
125 mM MgCl2

To make Oligo-dNTP mix (4.45 ml), mix the following
3.7 µl              2M HEPES, pH 6.6 (store at -20oC)
22.25 µl           12.5 M (98%) b-mercaptoethanol
3.99 mg           random hexamers
5.56 µl             100 mM dCTP
5.56 µl             100 mM dGTP
5.56 µl             100 mM dTTP
711 µl              H2O

To make Tris/MgCl2 mix (5ml), mix the following
3.125 ml          1M Tris-HCl pH 8.0
0.127 g            MgCl2 (MW 203.3)
1.875 ml          H2O

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