DNA/RNA QC : Agilent Bioanalyzer / TapeStation

The Agilent 2100 Bioanalyzer is a microfluidics-based platform for sizing, quantification and quality control of DNA and RNA. The platform is the industry standard for RNA sample QC and has mostly replaced labor-intensive gel electrophoresis.

 

DNA/RNA QC : Agilent Bioanalyzer / TapeStation

Platform
Agilent Bioanalyzer and TapeStation
Category
DNA and RNA QC
Primary Contact

Christopher Krebs, Manager, UM DNA Sequencing Core

ckrebs@umich.edu

734-763-8891

Typical Cost Click here to view internal pricing list!
Click here to view external pricing list!
Typical Output

You will get back a PDF report that contains the samples electropherogram and pseudo-gel images, as well as a concentration and, for RNA, a RIN value.

How to Request

Bring your samples to 2568 MSRB II or the DNA Sequencing Core in Building 14, NCRC where you will be asked to fill in contact and billing (shortcode) information on an electronic submission form.

Sample Requirements

We request a minimum of 5ul of each sample for analysis by Bioanalyzer. The samples for “QC only” should be in a separate tube from the portion of sample intended for other services within the core. This assists us in minimizing freeze thaw cycles.

Typical Turnaround

24 to 48 hrs

Critical Information

Bioanalyzer

RNA 6000 Nano Kit: 5-500 ng/ul, RNA Size range: 25-6000 nt

RNA 6000 Pico Kit: 50-5000 pg/ul RNA Size range: 25-6000 nt

RNA small RNA Kit: 1-100 ng/ul RNA Size range: 6-150 nt

DNA 1000 Kit: 0.1-50 ng/ul DNA Size range: 25-1000 bp

DNA 12K Kit: 0.5-50 ng/ul DNA Size range: 100-12000 bp

DNA High Sensitivity Kit: 5-500 pg/ul DNA Size range: 50-7000 bp

TapeStation

RNA ScreenTape: 25-500 ng/ul, RNA Size range: 100-6000 nt

High Sensitivity RNA ScreenTape: 1-25 ng/ul RNA Size range: 100-6000 nt

DNA 1K ScreenTape: 0.1-50 ng/ul DNA Size range: 35-1000 bp

High Sensitivity DNA ScreenTape: 10-1000 pg/ul DNA Size range: 35-1000 bp

Genomic DNA ScreenTape: 5-300 ng/ul DNA Size range: 200-60,000 bp

More Information

The RIN, or RNA Integrity Number, is a method for determining RNA quality. RIN is calculated algorithmically by including the 28s/18s ratio, the region before the peaks, signal areas, and intensities. These calculations enable use of the whole graph to make RNA quality assessments. We make an initial assessment of total RNA QC based on 260/280 and 260/230 ratios. However, we place the highest reliance for QC on the Bioanalyzer integrity assessment. Each sample is assigned a RIN score: 10 corresponds to a pure, un-degraded total RNA sample while a RIN of 1 corresponds to a completely degraded sample. The core recommends only using samples with RIN's greater than 7.

Link to additional information for interpreting Agilent 2100 Bioanalyzer data: http://www.urmc.rochester.edu/fgc/documents/Bioanalyzer_Interpretations_forcustomers.pdf