Sample Amount Required
- For each sample, we ask that you try to send in at least 1-3 µg of total RNA. Please transfer your sample to a 1.5ml microcentrifuge tube for shipment (0.5 ml or smaller PCR tubes can crack when frozen).
Total RNA Extraction
- It is very important to use a total RNA preparation procedure that does not remove the low molecular weight RNA fractions.
- Make sure to use one of the commercially available RNA extraction kits that are specifically developed for small RNA studies, such as Norgen Biotek, Qiagen miRNeasy, Ambion miRVana, etc. Please refer to corresponding manufacturer’s manuals. Use only one type of extraction kit for all the samples of your project.
- Many laboratories have obtained excellent results from total RNA samples extracted using Trizol methods. However, skills, experiences and sometime sample types may become critical factors in obtaining consistently good sample qualities. According to our statistics, the method has an overall higher failure rate than column-based commercial methods, although the rate varies among different laboratories. If you must use a Trizol method, we recommend modifying precipitation step by doubling the usual isopropanol volume and leaving the RNA at -80°C for 10-20 minutes so as to ensure the precipitation of small nucleic acids. Some laboratories perform the precipitation step twice and/or perform a post-precipitation wash twice in order to clean up the sample. You will need to perform some tests in order to find a proper protocol for your sample.
- Please transfer your sample to a 1.5ml microcentrifuge tube for shipment (smaller tubes can crack when frozen).
- International Orders – for those customers shipping samples oversees, please download a copy of our International Shipping Instructions.
- Note – We now offer an RNA extraction service for our US customers who would prefer to just send us cells or tissue samples.
Quality Control of RNA Sample
- A quality control check of your total RNA sample is included in our “Total RNA to Data” comprehensive service.
- If desired, you can check the RNA quality with a Bioanalyzer, or a 1-1.5% agarose gel. High quality RNA will show a 28S rRNA band at 4.5kb that should be twice the intensity of the 18S rRNA band at 1.9kb. Excessive smearing indicates degraded RNA.
- It is important to check the UV spectrum of your sample with a spectrophotometer. The 260nm/230nm ratio should be greater than 1.0 and the 260nm/280nm ratio should be above 1.8.
- Note – We will notify you if samples fail our quality control analysis before proceeding with your microarray experiment. There will be a small processing charge for samples that fail quality control.
Placing Your Order
- Please see the ordering page for instructions on placing your small RNA sequencing order.