Thank you for nice presentation. I am interested in the varian mRNAs of the ERalpha.
(1) My first question is technical one about semi-quantitative PCR. It was described that PCR was done in 30 cycles. I am afraid that PCR products might saturate after 30 cycles of amplification and it might interfere the semi-quantitative analysis even through triple primer PCR. Have you checked other cycles of amplification, for example 25 cycles?
(2) During the PCR, shorter fragments would be amplified preferentially than longer ones. Therefore I am afraid that the initial amount of ERD3 and ERD5 might be much smaller than wild type transcript. I should like to know your interpretation of the physiological significance of such minor transcript.
(3) In contrast, amplified fragments of ERC4 is much longer than those of wild type transcript. I should like to know the sequence and the structure of this transcript in detail.
Tue Dec 15