The Agilent Tapestation provides an automated alternative to traditional gel electrophoresis, allowing researchers to analyze the quantity and size of DNA or RNA samples from only a few microliters.
The Genomics Core can process samples on the TapeStation in three different configurations. First, RNA samples can be run on an RNA specific tape, resulting in the calculation of a RIN (RNA Integrity Number) score for each sample. The RIN score provides a useful metric of the RNA quality; for example, we ask that samples have a RIN score of at least 6.0 and preferably 7.0 before choosing a ribodepletion or poly(A) selection protocol, while our SPIA total RNA protocol can reliably generate cDNA from samples with RIN scores as low as 2.0.
The three images to the left are representative of the spectrum of RNA quality and show the output electropherogram provided by the TapeStation for an RNA sample. The top image, with poorly-defined 18S and 28s peaks, is from a sample with a RIN score of just 3.1; the middle image is of a sample with a RIN score of 6.1; and the bottom image, with the sharpest peaks and least background, is from a sample with a RIN score of 9.8.
The second two options for the TapeStation are both for DNA; the HSD1000 tapes are ideal for samples with low concentration and small size, while the D5000 tapes are better for samples with higher concentrations and longer lengths. On the right, the top image is of a sample run on a D5000 tape, and the bottom image is from a HSD1000 tape.
No matter what tape is used, each electropherogram displays the sample peak as well as marker peaks (one peak for RNA tapes, and two for DNA tapes). These marker peaks normalize each sample to the ladder, which is run independently, and enable accurate quantification and fragment length calculation.
The TapeStation in the Genomics Facility is open for researchers to use following training on the instrument, or technicians can process samples on your behalf. Please visit our iLab page to reserve instrument time or make a request for samples to be run.
- DNA, RNA and next generation sequencing quality control
- File16.52 MB