An example of the RT-PCR analysis performed on the non-lethal gene trapped lines is shown below. Primers complementary to exons flanking the insertion site are used in an RT-PCR reaction to determine whether the gene trapping vector has effectively disrupted transcription of the endogenous message in homozygous mice.
Three samples were run for each tissue type under the following conditions:
Tissue in the absence of gene specific reverse transcript (RT) and Actin control primers.
Tissue in the presence of RT and the absence of Actin primers.
Tissue in the presence of RT and Actin primers.