( s: The Northern blot contai...)Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells

s: The Northern blot containing total RNA from CHO cells was cut into duplicate sections, and each was prehybridized in 3 ml of Stratagenes QuikHyb hybridization solution for 15 minutes in Stratagenes PersonalHyb hybridization oven, according to the QuikHyb manuals instructions. Each blot was hybridized with 3 x 10⁶ cpm of one of the probes (10⁶ cpm/ml) for 1 hour, also according to the manual. The membranes were washed twice for 10 minutes in 2X SSC/0.1% SDS at room temperature and twice for 15 minutes with 0.1X SSC/0.1% SDS at 60C. The membranes were enveloped in plastic wrap and exposed to Kodak XAR-5 film at -80C (with an intensifying screen) for varying amounts of time. An overnight exposure detected the low to moderate-abundance MSH2 transcript from as little as 200 to 500 ng of total RNA. After a 4-day exposure, the expected 3.1-kb band was observed from as little as 100 ng of total RNA, with a minimum of background (Figure 2).

Fig.2

REFERENCES

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3. Dolter, K. and Braman, J. (1999) Strategies 12: 3.

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8. Felts, K., et al. (1992) Strategies 5: 3738.

9. Jerpseth, B
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