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Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells


Integrated system simplifies and improves Northern blot analysis

Karen Dolter Jeff Braman
Stratagene

Streamline RNA analysis with a whole host of products from Stratagene. First, the StrataPrep total RNA miniprep kit is used to isolate ample yields of high-quality RNA from tissues and cells, then the RNA is ready to use in a number of applications, including Northern blot analysis. This procedure features the following products: the Joule Box minigel electrophoresis system, the Stratalinker UV crosslinker,* the prostar first strand RT-PCR kit, Stratagenes primer sets for RT-PCR, the StrataPrep PCR purification kit or the StrataPrep DNA gel extraction kit, the Prime-It II random primer labeling kit, NucTrap probe purification columns,** QuikHyb hybridization solution, and the PersonalHyb hybridization oven. When used together for Northern blot analysis, this excellent integrated system permits high sensitivity and reproducible results.

Northern blot analysis of gene transcription is dependent on such factors as isolating intact, high-quality RNA, probe-labeling to high specific activity, removing unincorporated radiolabeled nucleotides, and hybridizing with conditions that maximize the signal-to-noise ratio. Products developed at Stratagene eliminate many variables that lead to poor quality Northern blot results. Consequently, we used these products for a Northern blot analysis of MSH2 gene transcription in a hamster cell model system.

MSH2 is important for DNA mismatch repair, and mutations in this gene are responsible for many cases of hereditary nonpolyposis colorectal cancer.1 The MSH2 gene encodes a 3.1-kb mRNA that is expressed at low to moderate levels.2 The larger-than-average size and
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