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Simple Isolation of RNA from Tissue and Cultured Cells

2.0 0.22

2.2 0.11

4

Liver

4.2 1.05

2.1 0.02

4

Lung

1.2 0.15

2.1 0.04

2

Spleen

3.6 0.88

2.2 0.19

4

Testis

1.6 0.36

2.2 0.14

4

Assay for DNA Contamination

With the RT-PCR technique, small quantities of genomic DNA contamination can be detected in RNA samples. This method was used to demonstrate the effective removal of DNA using the StrataPrep total RNA miniprep kit (Figure 3). Primers specific for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) sequences (present in approximately 200 copies in mouse and rat genomes2) were used, providing an especially sensitive assay for the presence of DNA. PCR amplification of GAPDH sequences from cDNA shows the expected 0.2-kb fragment (Figure 3: Lanes 1, 3, 5, and 7). When the DNase treatment step is omitted from the protocol and reverse transcriptase is omitted from the cDNA synthesis reaction, the presence of DNA is detected in the PCR (Figure 3: Lanes 4 and 8). However, when the StrataPrep total RNA miniprep protocol is car
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