Detection of Rab27a mRNA. (Panel A) Schematic depiction of transgenic (rat) versus endogenous (mouse) Rab27a coding sequence. EcoRI or SmaI restriction sites as well as resulting fragment sizes are indicated. (Panel B) Comparison of transgenic versus endogenous Rab27a expression in the eye by RT-PCR and restriction digestion of PCR products. Rab27a mRNA from wild-type and independent mutant transgenic lines (Rab27aT23N and Rab27aN133I) amplified by RT-PCR was digested with EcoRI or SmaI and electrophoresed as described under "Methods". Rat Rab27a cDNA was used as a control. (Panel C) Determination of Rab27a expression (transgenic and endogenous) in eyes by RT-PCR (+). The oligonucleotides and conditions were as described under "Methods". The PCR products were analysed on an agarose gel stained with ethidium bromide. Reactions performed without reverse transcriptase are indicated (-). The arrows on the left-hand side indicate the positions of DNA marker sizes. Hprt expression was used as an internal control.
Ramalho et al. BMC Cell Biology 2002 3:26 doi:10.1186/1471-2121-3-26