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Simple Isolation of RNA from Tissue and Cultured Cells

ried out according to the kits instructions, no DNA contamination is detected (Figure 3: Lanes 2 and 6).

Fig.3

Detecting RNA of Varying Abundance

RNA isolated with the StrataPrep total RNA miniprep kit was used to synthesize cDNA, which served as a template for amplification using Stratagenes RT-PCR Primers for Human Gene Transcripts of Varying Abundance Levels,3 according to the instructions provided. The presence of PCR products for high- (-actin), medium- (g-actin) and low- (protein phosphatase 1, ADP ribosylation factors 1 and 3, and ornithine decarboxylase) abundance mRNA shows that a wide spectrum of RNA molecules is present in the sample (Figure 4).

Fig.4

Sensitive Detection of RNA Using Molecular Beacons

Molecular beacon technology provides an elegant and efficient method for PCR and RT-PCR that does not require gel electrophoresis of the products and permits accurate quantitation.4 The Mx4000 molecular beacon expression analysis kit for mouse -actin5 was used for real-time RT-PCR analysis of RNA isolated from NIH/3T3 cells with the StrataPrep total RNA miniprep kit. RNA is readily detected from a sample as small as 1 pg in a target titration of 1 100,000 pg of total RNA (Figure 5). These results demonstrate that the RNA isolated using the StrataPrep kit is of the high quality required for real-time RT-PCR.

Fig.5

Other Spin-Cup-Based Total RNA Isolation Kits

Most spin-cup-based total RNA isolation kits from other manufacturers do no
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