( place by a plastic microfuge rack. UV l...)Inactivation of Contaminating Templates in PCR Using High Intensity Ultraviolet Light

place by a plastic microfuge rack. UV light was delivered by the GS Gene Linker chamber for 30 minutes. After UV irradiation was initiated, both a capped and an open tube were removed every 10 minutes from each of the lambda and β-actin samples. Control tubes for the lambda and β-actin templates consisted of untreated tubes that were not irradiated (0 seconds).

After UV irradiation, an aliquot from a master solution mix, containing AmpliTaq enzyme, appropriate primers, reaction buffer, and dNTPs was added to each tube. The tubes were then subjected to 30 cycles of PCR and analyzed on a 3% NuSieve 3:1 gel.

Results
Amplification of the lambda genomic DNA results in a 500 bp fragment, which can be seen in the control lane (Figure 1, lane 1), and amplification of the β-actin product results in a 268 bp fragment which can also be seen in the control lane (Figure 2, lane 1). With increased levels of UV irradiation, the amount of detectable PCR product decreases, as visualized by ethidium bromide staining. After 30 minutes of UV irradiation, both of the templates have been inactivated. As demonstrated in Figure 1 and 2, UV irradiation is an effective means of inactivating templates, whether the tubes are capped or closed. However, our data suggest that capping the tube and placing it on its side in the middle of the chamber is more effective than leaving the tube open in an upright position.

For best results, we suggest the following procedure:
Cap the desired number of microfuge tubes
Place the tubes in the middle of the GS Gene Linker oven.
Select 900 seconds (15 minutes) and press start.

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