Homogenizing your samples:
We weigh and transfer your samples on dry ice to specialized tubes with an o-ring to prevent any sample loss. Each sample will be subjected to homogenization via bead mill in short intervals to prevent heat degradation.

RNA extraction:
RNA is then immediately extracted and tested for quality and quantity utilizing a nanodrop spectrophotometer, we will send you the report generated for each sample. In the case of RNA being low quality we will run a clean-up step, free of charge, and retest on the nanodrop spectrophotometer. We ensure the RNA has an A260/A280 ratio of 1.8-2.1 and an A260/A230 ratio of 2.0-2.2. 

cDNA synthesis:
One microgram of extracted RNA is immediately converted to cDNA. DNase incubation is performed to remove genomic DNA prior to cDNA synthesis.

RT-PCR:
Real-time PCR assay for any gene of interest + 2 housekeeping genes that are valid for the specific sample type / tissue type being assayed (we can select housekeepers for you)
These are done in a 3 color system within the same well. A fourth color containing ROX loading dye is also assayed. Our set up aims to give reproducible, robust data in a single well.

Results:
We will issue you with an excel file with cycle threshold results, melt curve data and include statistical comparisons between sample groups with delta-delta-CT values calculated based on the geometric mean of the 2 housekeeping genes. We will also send you quality control data demonstrating that your assay is working on a serial dilution of your cDNA so you can rest assured that this assay was performed within the quantitative range for your gene targets and housekeeping genes in your sample.  We also standardly include example text to use in publications for RT-PCR methods, statistical analysis, figure legends and results sections.

Return shipment of your samples:
For $120 we will ship excess samples, RNA and cDNA back to your lab
For $145 we will also include your PCR product, transferred from the well plate into tubes if required

Sample notes:
We accept samples frozen sent on dry ice (recommended) or in RNAlater.
Contact orders@genewavepcr.com if your samples are BSL3