To enable researchers at the Royan Stem Cell Institute to access molecular data for validating cellular assays, a centralized molecular biology laboratory facility was established.

The fundamental differences between various cell types—such as neurons, hepatocytes, osteocytes, or blood cells in an individual—do not stem from variations in genomic DNA sequences but rather from the differential expression of tissue-specific genes during development. One of the key stages in gene expression regulation occurs at the level of mRNA synthesis, which is measured using PCR. While the Central Laboratory provides polymerase chain reaction (PCR) facilities, the technique that allows researchers to quantify low-abundance mRNA—mostly obtained from tissue samples and cultured cells—is real-time PCR, which is the core function of this laboratory.

Currently, the Molecular Biology Laboratory is involved in executing over 40 research projects. In general, the services and activities of this lab include:

• Primer Design and Storage: Verification of primer sequences and storage in the lab’s primer bank, which contains over 800 pairs of human primers and 600 pairs of primers from other species.

• RNA and DNA Extraction: Isolation from cultured cells or tissue samples, followed by assessment of RNA integrity and purity for RT-PCR or microarray analysis.

• cDNA Synthesis (via reverse transcription).

• Gel Electrophoresis and Staining: Facilities for ethidium bromide, GEL RED, and SYBR GREEN staining.

• Real-time PCR (using ABI7500 Applied Biosystem and Rotor Gene 6000 Corbett systems).

• Real-time PCR Data Analysis.

• Isolation and Analysis of Regulatory Sequences of genes involved in stem cell differentiation.

• Gene Cloning for functional studies in the lab.

• Mycoplasma Contamination Testing using PCR.