FIG. 6.

Allelic expression analysis of the human 91H RNA. Strand-specific reverse transcription was performed on total RNA from T47D cells with a reverse primer for H19-specific RT and a forward primer for 91H-specific RT (see the Materials and Methods). PCR were performed with primers located on each side of the AluI polymorphic restriction site as indicated in the figure (HP1 and HP2). The PCR products were then digested with AluI and separated into agarose gel. Note that, as expected, the size of the H19 amplification fragment is smaller than those obtained for 91H RNA amplification because of the removal of the 81-bp H19 intron 5 in the H19 RNA. ND, not digested; D, AluI digestion; -RT, amplifications on control reactions made without reverse transcription. gDNA, control amplification on T47D cell gDNA.