全自动电泳疑问解答FAQs

Analysis Kits (Chips and Reagents)

Experion Automated Electrophoresis — Sample Types

What types of samples can the Experion separate and analyze?
The Experion automated electrophoresis system separates and analyzes protein and RNA (either mRNA or total RNA).

Experion Automated Electrophoresis — Analysis Kit Ordering Options

What are the various analysis kits available for the Experion system? What ordering configurations are available for these analysis kits?
The Experion system contains three analysis kits: The Pro260 kit for protein separation and analysis, the RNA StdSens kit for standard-sensitivity RNA detection, and the RNA HighSens kit for high-sensitivity RNA detection. All three analysis kits are available as 10- or 25-chip kits, for a total of 6 ordering options.

Experion Automated Electrophoresis — Consumables

What consumables will be sold separately for the Experion system?
Experion Protein and RNA chips (10-count) will be sold separately, as will electrode cleaning chips and reagents, DEPC-treated water, and spin filters.

Experion Automated Electrophoresis — Dynamic Range of Detection

What is the dynamic range of detection for the Experion system? What concentrations of proteins can be separated and detected with the Experion Pro260 analysis kit? What levels of RNA can be detected with the RNA HighSens and StdSens analysis kits?
The Experion Pro260 analysis kit separates and detects 2.5–2,000 ng/µl protein. The RNA HighSens analysis kit is designed for high-sensitivity RNA separation and detection down to 200–5,000 pg/µl RNA, and the RNA StdSens analysis kit detects 25–500 ng/µl RNA.

Experion Automated Electrophoresis — Sensitivity of Detection

What are the limits of protein and RNA detection with the Experion system?
The Experion system can detect 10 ng carbonic anhydrase (S/N > 3) with the Pro260 analysis kit, 5 ng/µl 18S rRNA in a total RNA sample (S/N > 3) with the RNA StdSens analysis kit, and 200 pg/µl 18S rRNA in a total RNA sample (S/N > 3) with the RNA HighSens analysis kit.

Experion Automated Electrophoresis — Sample Volumes

How much protein or RNA is required to run the Experion system? What sample volumes are required for the Experion Pro260, RNA HighSens, and RNA StdSens microfluidic chips?
The Experion chips require 4 µl of sample per well for protein analysis and 1 µl of sample per well for RNA analysis.

Experion Automated Electrophoresis — Minimum and Maximum Volumes in Chip Wells

What are the minimum and maximum volumes that the sample and gel wells on Experion chips can hold?
Sample wells can hold 4–10 µl (sample, ladder, and destain), and the gel well holds 7–18 µl gel matrix.

Experion Automated Electrophoresis — Shelf Life of Chips and Reagents

What is the shelf life of the chips and reagents in the Experion Pro260, RNA HighSens, and RNA StdSens analysis kits?
Reagents and chips of the Experion Pro260 analysis kit have a shelf life of 12 months. RNA chips have a shelf life of 15 months and the RNA reagents, 12 months.

Experion Automated Electrophoresis — Storage Temperature for Chips and Reagents

At what temperatures should the chips and reagents in the Experion Pro260, RNA HighSens, and RNA StdSens analysis kits be stored?
Protein and RNA chips may be stored at ambient temperature. Reagents should be stored at 4°C.

Electrophoresis, Priming, and Vortex Stations

Experion Automated Electrophoresis — Time Required for Chip Priming

How long does it take to prime a chip for the Experion system?
After initial setup, priming of the gel matrix takes 1–1.5 min in the automated priming station.

Experion Automated Electrophoresis — Number of Samples per Chip

How many samples can be run per chip on the Experion system?
The Experion Pro260 chips can accommodate 10 protein samples and the RNA StdSens and HighSens chips can accommodate 12 or 11 RNA samples, respectively.

Experion Automated Electrophoresis — Run Time

How long does it take to run a chip on the Experion system?
Chips that are completely loaded with sample generally require 30 min per run. Less time is required for fewer samples.

Experion Automated Electrophoresis — Automated Steps

What steps of the electrophoresis process does the Experion system automate?
The Experion system combines electrophoresis, staining, destaining, imaging, band detection, and basic data analysis into a single, automated, 30 min step.

Experion Automated Electrophoresis — Detection Method

How are samples labeled in the Experion system? How are they detected?
In the Experion system, protein and RNA samples are labeled with a dye and are detected by laser-induced fluorescence.

Experion Automated Electrophoresis — Sample Labeling Process and Molecular Weight Determination

In the Experion system, how are protein and RNA samples labeled with dye? Does this labeling process affect the apparent size of proteins and protein MW determinations?
The Experion dye intercalates into the protein or RNA molecules by a process that does not appear to influence the migration patterns or apparent MW of proteins. Proteins in the Experion Pro260 ladder also bind the dye, so any effect is taken into account through normalization to the protein ladder.

Experion Automated Electrophoresis — Additional Equipment Required

What additional equipment/supplies are required to run the Experion system, aside from the Experion instrument and analysis kit?
Additional equipment require and sold by Bio-Rad are: the priming station for introducing gel into the chip, and the vortexer, which is required for all RNA applications. A computer with a USB port and cable and a 10 µl pipet are also required and must be supplied by the customer.

Software

Experion Automated Electrophoresis — Software Calculations

What calculations are performed automatically by Experion software?
For protein samples, Experion software automatically calculates MW, sample concentration (µg/ml), % of total sample, standard deviation, mean value, and % CV. For RNA samples, Experion software automatically calculates the total RNA concentration (ng/ml), mRNA concentration (ng/ml), rRNA contamination, 28S/18S or prokaryotic (16S) ratio, standard deviation, mean value, and % CV.

Experion Automated Electrophoresis — Data Display, General

How does Experion software display separation data?
Experion software displays data in a conventional electropherogram, or peak view, and in a simulated gel view.

Experion Automated Electrophoresis — Data Display, Simulated Gel View

What is the simulated gel view that is displayed by Experion software?
Experion software detects each protein or RNA moiety as it passes through the detector and creates a virtual gel image based on the migration of the protein or RNA relative to the standard.

Experion Automated Electrophoresis — Data Display, Results of Calculations

How are the results of calculations displayed by Experion software?
Calculations are presented in two different formats: the result table organizes data to compare information well-to-well, and the protein/RNA table organizes data to compare information (concentration/statistics/% total sample) of a particular protein or RNA of interest from the different wells.