Agilent Bioanalyzer 2100
The Agilent bioanalyzer utilizes a microchannel based electrophoretic cell that allows rapid and sensitive investigation of nucleic acid samples. It is ideal for looking at very small amounts of RNA in order to assess integrity, and to estimate quantity, before labeling. Thus it is used routinely in microarray analyses to ensure quality control of input experimental material. The instrument is available to investigators to analyze nucleic acid samples for these and other purposes. Different fluidic cells, or "chips," are used depending on the type of analyses desired and amount of material present. These are purchased through Agilent; more information can be found at the Agilent bioanalyzer site. The facility will maintain a stock of RNA Nanochips only, and the accessory RNA ladder (used for sizing and quantification). Users can purchase these materials from the facility, or they can provide their own. In addition to the consumables charge, there will be an annual fee for training on the instrument; scans can be done at no charge but documention of use on a sign up list will be required. Output is most conveniently converted to a pdf file then transferred to a flash drive/memory stick. Several typical examples of total RNA profiles run on the bioanalyzer can be found at this Ambion hosted web site.
You must have a Genome Center account to use the bioanalyzer, which you can create here if necessary.
If you have never used the Bioanalyzer, you must arrange a time with us to be trained. Training is always carried out in conjunction with an actual run, so it's wise to have all samples that fit in a particular chip (up to 12) ready to go. Facility instruments that require user training involve training with facility personnel, not fellow labmates. There is a limit of four people during a training session and two separate training sessions can be done for the yearly fee. For more information on training, please look over our "Equipment" web page.
If you need the RNA NanoChip materials from our facility call or email us first to make sure someone will be available to provide them. Once a confirmed day and time has been established with our facility (for training and/or RNA NanoChip materials) follow the directions below for reserving instrument time. Finally, we strongly recommend familiarizing yourself with the bioanalyzer troubleshooting and maintenance manual. Not only will it help with your initial training, but it will give you a more detailed overview of the functions and capabilities of the instrument beyond what we cover. This manual is a 4.7 Mb pdf download.
If you have already completed a formal training with facility personnel and have your own materials, please use the on-line calendar to reserve time during normal working hours at least 24 hours ahead of time to ensure availability. Put your name in the "subject" box, and phone number and laboratory PI in the "description" box. If your lab has a particular DaFIS # to be used for equipment use, please indicate this as well. To delete a scheduled calendar date, contact Charlie or Heather.
A condensed instruction sheet summarizing the protocol is available and should be consulted before doing your run; among other things this document will explain what you need to provide and what we have here.
The standard fee for bioanalyzer use includes the 12 sample RNA Nanoassay chip, the gel matrix and associated dyes and buffers, and the RNA molecular weight ladder. You will need to bring some additional supplies for the procedure:
1. Pipettors and tips to transfer 0.5-10 ul volumes
2. Gloves
3. We recommend bringing your samples over as 2 uL aliquots in 0.5 mL tubes for the denaturing step. For the Nano chips, RNA should be in the concentration range of 5-500 ng/ul. You can use available inserts to quick spin these in a microfuge following denaturation.
4. You are responsible for keeping your samples cold if desired. Also, a quick chill step is recommended following denaturation.
5. Flash drive to copy results onto. If you don't have a flash drive, results can be emailed to you.