Progress: RNA Selection of S100A4

The full proposal has been posted in Dropbox, the abstract is here.

Progress, Results, and Discussion
One round of bead based selection with N34 RNA against s100A4 have been completed. The gel image of the ccPCR can be found below, of W0, W3, and E1 from selection. Large scale PCR was performed at 12 cycles for R1 and transcription was performed. The PAGE after transcription showed a band that was unusually below the gel front and a crash soak was performed to determine RNA concentration. Concentration was low and therefore transcription will be repeated for R1.

Fig 1: ccPCR R1 S100A4. Significant amplification

in elution cycles 12, 15, and 20. Optimal cycle is

seen in cycle 12 of elution.

Problems Encountered

The unusual PAGE may have been caused by an error in the PAGE or during transcription. Both will be redone before starting the next round.

Conclusion and Future Work

One round of bead based selection against S100A4 is almost completed. The concentration at the end of R1 was 24.6 uM and since a higher concentration would be preferable, transcription will be reperformed. After a successful round one, a negative selection will be performed to decrease nonbinding protiens and posted for the next progress report.