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RT-PCR Primers for the Study of Apoptosis

a specific band at the expected length: For ICE- b, a 600-bp band was observed; for CPP32, a 540-bp fragment was observed and for ICE-Lap-3, a 600-bp band was observed. The genomic DNA and mouse and rat cDNA did not generate similar PCR products. The relatively weak bands seen for CPP32 may reflect the abundance of the CPP32 cDNA present in the cDNA library. Increasing the concentration of template in the reaction by 50% did not result in a significant increase of PCR products.

Figure 3

The ICE-Lap-3 and Fas antigen primer sets did not amplify the expected PCR product from the human muscle cDNA library. Repeated PCR attempts generated the same results. This may be explained by the lack of specific cDNA target in the library. A control primer, human b-actin, was used to test the cDNA library. An expected band was amplified for all human cDNA libraries tested (data not shown).

Mouse Fas Antigen and Fas Ligand

Several different mouse cDNA library templates were used to amplify Fas antigen and Fas ligand gene products (Figure 4). Both mouse Fas antigen and mouse Fas ligand RT-PCR primer sets amplified PCR products of the expected sizes. The Fas ligand gene has a wide distribution range in different tissues; however, the Fas antigen gene is differentially expressed in certain tissues. No amplification occurred in the control rat and human cDNA libraries.

Figure 4

Rat Fas Antigen

The rat Fas antigen gene was amplified in rat brain, lung, and heart cDNA libraries (Figure 5). The rat heart cDNA library generated a PCR product larger in size than expected
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