The PMGC provides several services for assessing the quality and quantity of samples prior to undertaking further experiments.
Please click on an image below for more information.
|Qbit® 2.0 Fluorometer
|Infinite® 200 PRO NanoQuant
|Bioanalyzer kit||Amount requested||Concentration range|
|RNA Nano||2-3 µl total RNA in RNase-free water||100-200 ng/µl|
|RNA Pico||2-3 µl total RNA in RNase-free water||5-10 ng/µl|
If you do not know the concentration of your sample, you will be charged for running the sample even if the concentration is too low to detect. We kindly ask that you dilute concentrated samples accordingly as high concentrations can cause problems with the assay.
The Protein 230 and Protein 80 kits require 4 µl of sample at a concentration between 0.1-2 µg/µl. The High Sensitivity 250 kit requires 4 µl of sample with a concentration >10 pg/µl, which is superior to silver stain SDS-PAGE.
The Agilent Bioanalyzer can also be used to measure mRNA, amplified RNA, labelled-cDNA, and other nucleic acids, however, the standard service we are offering currently applies to total RNA only. If you have specific needs, please contact us.
Assessing the quality of your RNA sample before proceeding with labelling, etc., can save a lot of time and money.
It can also reduce the time spent troubleshooting if something goes wrong at the next step.
If you know your RNA was good, you can rule out degraded RNA as the cause of your problem.
The Agilent Bioanalyzer is a convenient replacement for standard denaturing agarose gels; it is much faster, requires only a small amount of sample, and is more environmentally-friendly. The Bioanalyzer uses capillary electrophoresis (lab-on-a-chip technology) to move the sample through a gel matrix. An intercalating dye in the matrix allows the nucleic acid to be detected as it moves through the capillary. The electropherogram provides the user with a detailed trace of each sample.
For a list of compatible buffers, please visit Agilent's site which list all known compatible buffers.
Yes, the Bioanalyzer is able to analyze proteins under reducing (in the presence of DTT) and non-reducing conditions. All assays are run under denaturing conditions.
The RNA Integrity Number (RIN) is a software tool that enables users to estimate the integrity of total RNA samples. The RIN is a numerical assessment of the integrity of RNA and allows for the direct comparison of RNA samples. For more information, please visit Agilent's RIN web page.
|High quality total RNA
The electropherogram should have 2 distinct peaks around 41 and 47 seconds (corresponding to the 18S and 28S RNA), as well as the marker around 22 seconds, and the baseline should be flat.
|Partially degraded total RNA
The presence of a broad peak between 23-30 seconds indicates the presence of small (degraded) RNA products and tRNA. When RNA is partially degraded, the 18S and 28S peaks are often not very distinct and the baseline is raised.
|Degraded total RNA
Total RNA is completely degraded when distinct 18S and 28S peaks are not visible on the electropherogram and the baseline is raised. A flat baseline with no peaks may indicate that too little RNA was loaded for the assay and so is undetectable.