Electrophoresis of DNA or RNA on chips is performed with the equipment Bioanalyzer 2100 (Agilent). It is a fast and reliable method to determine the quality, integrity and concentration of such samples. The chips contain a series of interconnected fluidic microchannels that are filled with a fluorescent matrix. DNA or RNA is separated by electrophoresis according to size and visualised by fluorescence emission. The data is displayed as a fluorescence electropherogram and also as a gel-like image.
| Chip type | Separation range (nt) | Total DNA/RNA concentration required | Chip capacity (samples) | Parameters obtained |
|---|---|---|---|---|
| DNA 1000 | 25-100 | 0.5-50 ng/μl | 12 | concentration |
| HS DNA | 50-7000 | 5-500 pg/μl | 11 | concentration |
| Nano RNA | - | 50-500 pg/μl | 12 | RIN/concentration |
| Pico RNA | - | 0.5-5 pg/μl | 11 | RIN/concentration |
| Small RNA | <150 | 10-100 pg/μl (1) | 11 | %miRNA |
(1) RNA preparations enriched in smallRNA, the upper limit of concentration to be analysed shall be 20 ng/μl. The quantitative range of the test is between 50-2000pg/μl of miRNA. For highly enriched samples it is recommended to analyse different dilutions to ensure the measurement is within the quantitative range of the test.
Samples requirements
- Only 1 μl is needed for the analysis, although at least 4 μl must be sent. If the concentration of the sample is not known, it shall be determined by measurement of the absorbance in Nanodrop. In this case at least 5 μl of sample is required.
- If there is insufficient quantity of DNA or RNA measured by Nanodrop, the quality of the DNA or RNA will not be studied.
- Please send samples in 0.5 ml tubes, NOT in PCR tubes.
- Samples should preferably be sent in nuclease-free water. If samples are not in water, indicate the buffer used for elution or resuspension. A high concentration of salts may inhibit the electrophoretic mobility of the sample in the microchannels.
Delivery of results
- Results will generally be delivered within a maximum of 5 working days from receipt of the sample.
- The results shall be delivered as a PDF file including the electropherogram, a gel image and a table indicating the peaks identified, their size and concentration. Also, where appropriate, the RIN or miRNA percentage shall be indicated.
- After sending the results, the excess samples will be kept for a maximum of 15 days and then disposed of. If they are to be retrieved, please indicate this to the laboratory staff when sending them.
