Sample Preparation
Sample Amount Required
- For each assay, we ask that you try to send in at least 5 µg of total RNA. If you are limited in sample amount, please give us a call. We can accommodate smaller sample sizes. Please transfer your sample to a 1.5ml microcentrifuge tube for shipment (0.5 ml or smaller PCR tubes can crack when frozen).
- We prefer that you send total RNA but if you have already done the enrichment/purification for microRNA, that is OK. We ask that you try to send in at least 500 ng of enriched microRNA.
- If you have difficulty in obtaining the above quantities, please talk to our customer support. We may accept certain smaller quantities on a trial basis. In this case, we will prepare your samples and measure the processed RNA quantity before hybridization. If not sufficient amount is produced from the preparation, we will notify you to send in additional materials.
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 microRNA 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. - Note – If your sample tissue is prepared as FFPE (formalin fixed paraffin embedded), you must use an extraction kit designed specifically for FFPE samples. Standard RNA extraction kits will not work properly. Please see the accompanying application note.
- Note – If your sample is blood or plasma, please see the accompanying application note.
- There is no need to perform a small RNA enrichment step. We can accept fractionated microRNA but in this case, certain controls can not be included in your experiment.
- 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.
- Additionally, you can 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. (See exception for blood/plasma samples)
- 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
Determine Array Content and Select Additional Services
- See the list of available arrays and select the species or species group you are interested in.
- Select any additional services (such as rush service or in-depth data analysis).
- If you are placing a new order with the intention of normalizing this new data to previous data then the miRBase versions should match. This is particularly important for comparisons involving versions before 10.0 due to the significant changes in miRBase 10.0 and up.
Complete a Sample Submission Form
- Please complete a sample submission form prior to sending your sample to us. Email the form to us at:
sales@lcsciences.com. - A copy of the form should also be sent in the package along with your sample. (Note – please place the form in a separate waterproof bag from your samples.)
Packaging and Shipping
- Please transfer your sample to a 1.5ml microcentrifuge tube for shipment (0.5 ml or smaller PCR tubes can crack when frozen). Be sure the tube labels match those listed on your sample submission form.
- If you want to seal the tubes or hold them in a rack, please don’t use tape (it will crack when frozen), use parafilm.
- Pack the sample with dry ice in a thermal insulated shipping box.
- Ship the package by overnight carrier for delivery the next day.
- Note – We recommend to ship your samples Monday – Wednesday. Do not ship samples on Friday as they will sit over the week-end and deteriorate. Wait until the following Monday and ship for Tuesday delivery.
Mail the package to:
Attn: Sample Receiving
LC Sciences
2575 West Bellfort St., Suite 270
Houston, TX 77054
- International Customers – please download a copy of our International Shipping Instructions.
