Coincident In Vitro Analysis of DNA-PK-Dependent and -Independent Nonhomologous End Joining
Figure 4
Effects of reaction conditions on DNA end-joining activity. (a) Reaction buffer composition effects on HeLa WCE end-joining activity time course reactions in the presence of 5% PEG. Reactions were run at C for the times indicated in either of the two standard reaction buffers as described in the text. The mean values of triplicate reactions were plotted and the error bars indicate standard deviation. (b) Effects of MnCl2 on HeLa WCE DNA end-joining activity. Reactions were run with the addition of 0–5 mM MnCl2 to standard DNA end-joining reactions containing 5 mM MgCl2 for 2 hrs at C in the presence or absence of 5% PEG as indicated. (c) The DNA end-joining reactions employing MnCl2 (0–5 mM) in the presence of PEG were repeated with and without 10 M wortmannin (6 hrs at C). Reactions were run in triplicate and the error bars indicate standard deviation.
