• Created : 
  • Modified : 
  • Category :  Agilent Knowledge Portal
  • Entry Type :  Article
  • Product Type :  Automated Electrophoresis Systems
  • Component :  Bioanalyzer Systems TapeStation Systems
  • Product Name :  2100 Bioanalyzer Instrument 2200 TapeStation System 4150 TapeStation System 4200 TapeStation System
  • Task :  Operation Troubleshooting
Related Articles

  • Analysis of Yeast RNA on the Agilent Bioanalyzer

    This Information Applies To: 2100 Bioanalyzer, 2100 Expert software, RNA 6000 Nano kit (p/n 5067-1511), RNA 6000 Pico kit (p/n 5067-1513) Issue: Which assay should be selected in the Agilent 2100 Expert software when running yeast total RNA samples on the Agilent 2100 Bioanalyzer system? Resolution: The Bioanalyzer total eukaryotic RNA assays are validated for eukaryotic RNA samples with typical 18S and 28S ribosomal RNA (rRNA) peaks. Yeast is a eukaryote but has 18S and 25S rRNA peaks...

  • Intensity Loss for 200 nt Ladder Peak in the Agilent TapeStation RNA Assays

    This Information Applies To: 2200/4150/4200 TapeStation systems. RNA ScreenTape and High Sensitivity RNA ScreenTape assays,TapeStation analysis software Issue: The 200 nt peak of the Agilent RNA ScreenTape and High Sensitivity RNA ScreenTape Ladder has lost its normal intensity. An example of this phenomenon is shown in Figure 1 with the actual 200 nt peak highlighted with a red circle: Figure 1 . RNA Ladder with rudimentary 200 nt peak in red circle If the peak is below a certain...

  • Are TapeStation Assays Denaturing?

    This Information Applies To: Agilent 4200 and 4150 TapeStation Issue Are the Agilent DNA and RNA ScreenTape assays denaturing? Resolution The DNA ScreenTape assays in the TapeStation portfolio are not denaturing. Only the RNA and High Sensitivity RNA ScreenTape assays are denaturing. Components in the RNA and High Sensitivity RNA sample buffer partially denature the sample. There is also a heat denaturation step included in the RNA and High Sensitivity RNA operating procedure in the...

  • RIN Not Calculated or NA for Total RNA Samples in the Agilent Bioanalyzer 2100 Expert Software

    This Information Applies To: 2100 Bioanalyzer, 2100 Expert software, RIN NA, Error 4501, RNA 6000 Nano kit (p/n 5067–1511 ), RNA 6000 Pico kit (p/n 5067–1513 ) Issue: The RIN is not displayed in the software or shows as NA. If the RIN is NA, the sample will be flagged in red and error 4501 will be present with a description such as Unexpected signal... . Steps to follow: Check which assay was selected. A RIN, will only be calculated if a total RNA assay was selected. A RIN will not...

  • How to Determine the DV200 of FFPE RNA Samples on the Agilent TapeStation

    This Information Applies To: 4200, 4150 and 2200 TapeStation, TapeStation analysis software A02.02 or higher. Issue: When using the Agilent 4200, 4150 and 2200 TapeStation systems, the DV 200 of FFPE RNA samples can be calculated within the TapeStation analysis software. The DV 200 score is a quality score for evaluating quality of RNA derived from formalin-fixed paraffin-embedded (FFPE) samples established by Illumina Inc. in 2016. It represents the percentage of RNA fragments larger than...

Discrepancy Between Agilent RIN and RINᵉ Values

Answer

This Information Applies To: 2100 Bioanalyzer, 4150/4200/2200 TapeStation

Issue:

Discrepancy between Agilent Bioanalyzer RNA integrity number (RIN) and Agilent TapeStation RNA integrity number equivalent (RINe) values

Resolution: 

First, please read the following application notes about RIN and RINe:   

As can be seen in the application notes, in general, we expect RIN and RINe to correlate well.

However, if differences in RIN and RINe are observed, we have the following recommendations:

1. Ensure that the sample preparation protocol is followed exactly as recommended for each assay. For example:

  • For Bioanalyzer RNA assays, we recommend heat denaturing the samples before adding samples onto the chip. Although heat denaturation is optional to the workflow, it is highly recommended especially when comparing to RINe.  
  • For TapeStation RNA assays, we recommend heat denaturing samples after adding sample buffer. 

Therefore, it is important to ensure the same samples heat denatured once for a Bioanalyzer run are not used for a TapeStation run. This will cause the samples to be heat denatured twice, negatively affecting the RINe value. Also, for heat denaturation, proper tools including but not limited to properly sealable consumables and PCR cyclers.  

2. Ensure that ScreenTape and Bioanalyzer assays of comparable sensitivity and specifications are used when comparing data across the two platforms.

3. Ensure samples are loaded according to the listed specifications for each Bioanalyzer kit and TapeStation ScreenTape assay, and only compare the results of the same samples.

Finally, although we expect the two to correlate well, RIN and RINe use different algorithms. RIN uses a proprietary neural network-based algorithm to assess the entire electrophoretic trace. For RINe calculation, the upper ribosomal peak is not taken into consideration. Correct assignment of the lower marker is essential for RINe calculation whereas ladder is not required. In contrast, RINe represents the relative ratio of the signal in the fast zone to the 18S peak signal. Also, given that Bioanalyzer and TapeStation are two different platforms, differences in electrophoretic patterns as well as in RIN values compared to RINe values may occur depending upon samples. In this case, setting a separate threshold or quality cut off for RIN and RINe is recommended. 

For Research use only 

PR7000-7979

 Tip 

For further information please refer to

Agilent 2100 Bioanalyzer System Maintenance and Troubleshooting Guide
Agilent Information Center > Agilent TapeStation Systems > ScreenTape Assays > Good Measurement Practices
Agilent Information Centre > Agilent TapeStation Systems > ScreenTape Assays > Troubleshooting > Troubleshooting RNA Applications
Agilent Information Centre > Agilent TapeStation Systems > ScreenTape Assays > Troubleshooting > Frequently Asked Questions
 
Was this helpful?
Privacy Statement
Terms of Use
Contact Us
Site Help