Agilent Bioanalyzer and NanoDrop

Agilent Bioanalyzer

The Agilent 2100 Bioanalyzer is a microfluidics-based platform relying on the use of kits that include sample-specific reagents and chips. These kits provide a convenient method to determine sample integrity before a project gets underway. 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.

All projects submitted for Full Service are subject to Bioanalyzer QC testing. 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. Using the “Expert” software on board with the scanner, the Bioanalyzer assigns a RIN of 1-10 for each sample. A RIN of 10 corresponds to a pure, un-degraded sample. A RIN of 1 corresponds to a completely degraded sample.

Currently, Agilent manufactures two total RNA testing kits. Qualitative ranges for the Agilent 2100 Bioanalyzer RNA kits are as follows:

  • RNA 6000 Nano Kit: Range for RNA loaded = Total RNA: 5-500 ng/ul, mRNA: 25-250 ng/ul
  • RNA 6000 Pico Kit: Range for RNA loaded = Total RNA: 50-5000 pg/ul, mRNA: 250-5000 pg/ul

We request ~3ul of sample in the above concentrations for analysis by Bioanalyzer. The samples for QC should be in a separate tube from the portion of sample intended for RNA labeling. This assists us in minimizing freeze thaw cycles.

If you have RNA that you wish only to have tested for RNA quality, we will also perform RNA testing on the Bioanalyzer as a “stand alone” service.

 

NanoDrop 1000

The NanoDrop 1000 UV/Visible spectrophotometer makes measurements using a very small volume of sample generally without need for any dilution. Readings are reproducible and give accurate RNA and DNA concentration measurements. To make measurements 1ul of solution is required. Samples submitted to us lacking concentration information may be analyzed via the NanoDrop at additional cost to the project.

If you have RNA that you wish only to have tested for RNA concentration, we will also perform RNA testing on the NanoDrop as a “stand alone” service.