ATRA to the rescue!

We created the cDNA from the RNA because cDNA is far more stable and can be processed by our analytical tools.

The cDNA is checked for markers through qPCR. The first step of qPCR is to remove the tubes from the thermal cycler that has been creating the cDNA. We then lay out a “plate”. A plate can hold up to 324 samples, but can only test each sample for one gene marker. Since each test usually has 24 cDNA tubes, and each sample needs to be run in duplicate, so we can run up to 8 markers at a time. We are currently looking at our ectodermal cells and running markers to test for neural genes expression. We know that Retenoic Acid (also known as Vitamin A) is a posteriorizing signal. It acts by binding to DNA as a heterodimer at a RARE (retin

oic acid response element). This alters the configuration of the reinoic acid receptor which affects the binding of other proteins that induces and represses transcription of nearby genes. Until this point we have been using the small molecule TGF-Beta to agonize retinoid acid. Recently, advised by other lab members, we have experimented with other small retinoic acid molecule agonists. To our delight, we have found a new, far more effective molecule called ATRA. Our latest plate confirmed that ATRA has increased gene expression by more that fifteen fold! I’ll let you know about further lab developments in the future.