TapeStation Data Interpretation

  • Each lane contains a marker along with your sample. The marker is used to align the samples. The concentration and sizing is determined from the standard ladder loaded into lane one.
  • The 28S/18S ratio may be indicative of problems with the RNA, but not always. Even though the ideal ratio is 2.0, the TapeStation rarely reflects this ratio. If the ratio is greater than 2.0 it may indicate the presence of sheared single stranded genomic DNA which can run around the 28S band. If the ratio is less than 1.0, there are usually definite degradation problems with the RNA. More important than an absolute number is the visual sign of degradation.
  • The Agilent Expert software assigns a RINe1 number to each trace. It assigns a number according to how much signal is found between the 5S and 18S band, between the 18S and 28S bands, and after the 28S band. An RIN number of 10 is perfect score or excellent RNA. The software does not always call RINe numbers for prokaryotic RNA and the RINe can be misleading for samples containing additional RNA bands such as those from chloroplasts or a symbiotic RNA

Comparison of the Agilent 2100 Bioanalyzer and the 4200 TapeStation

 

1 See here for how the RINe compares to the 2100 Bioanalyzer RIN

Free software from Agilent is available to view your data on a PC.