Summary
The TapeStation system allows for automated electrophoresis of most DNA and RNA samples. Typical applications include quality control (QC) of NGS libraries and RNA extractions (ie. RIN). This method excels in the determination of fragment length distribution of nucleic acid samples, but is not typically relied on for accurate quantification. The TapeStation can be used in place of the BioAnalyzer for most applications, but cannot currently be used for small/micro RNA analysis.
Determining which test to select
The first thing to consider is what your sample type is: RNA, DNA or genomic material. For genomic DNA please select the genomic DNA service. For RNA and DNA there is a choice between high-sensitivity and not. High-sensitivity is only required when the sample concentration is too low to be run on the regular assay, typically lower than 25 ng/uL for the RNA assay and 1 ng/uL for DNA. For DNA product size also needs to be considered. If your product is under 1000 bp please select the D1000 assay (regular or high-sensitivity) and if it is between 1000 and 5000 bp please select the D5000 assay (regular or high-sensitivity). For anything larger than 5000 bp please select the gDNA assay.
On your order form please include which analysis should be done (DNA, genomic DNA, high-sensitivity DNA, RNA, high-sensitivity RNA).
If you are unsure about what assay to select please contact us to discuss your needs
Request Form: AAC TapeStation Request Form
Equipment Used: Agilent TapeStation 4150
|
Assay |
Set-up |
Per Sample |
|
RNA |
$10.50 |
$5.50 |
|
HS RNA |
$12.00 |
$6.25 |
|
D1000 |
$8.00 |
$5.50 |
|
HS D1000 |
$11.00 |
$7.75 |
|
D5000 |
$12.75 |
$5.25 |
|
HS D5000 |
$17.25 |
$7.75 |
|
gDNA |
$8.75 |
$8.75 |
Set-up fee is a flat fee charged per submission in addition to the per sample charge.
Set-up fee covers DNA/RNA ladders, technician time, and instrument overhead. Contact facility for more information about ladders and when to use them.
FAQs
What analysis should I use?
The first thing to consider is what your sample type is: RNA, DNA or genomic material. For genomic DNA, please select the genomic DNA service.
For RNA and DNA there is a choice between high-sensitivity and not. High-sensitivity is only required when the sample concentration is too low to be run on the regular assay, typically lower than 25 ng/µL for the RNA assay and XX for DNA.
For DNA, product size also needs to be considered:
- If your product is under 1000 bp please select the D1000 assay (regular or high-sensitivity)
- If it is between 1000 and 5000 bp please select the D5000 assay (regular or high-sensitivity)
- For anything larger than 5000 bp please select the gDNA assay
On your order form please include which analysis should be done (DNA, genomic DNA, high-sensitivity DNA, RNA, high-sensitivity RNA).
If you are unsure about what assay to select please contact us to discuss your needs.
How should I provide my samples?
| # of Samples | Recommended storage tube/plate |
|---|---|
| 1 to 8 | Individual tubes |
| 8 to 24 | 8 tube strips or partial 96 well PCR plate* |
| More than 24 | Partial or whole 96 well PCR plate* |
*if using a partial 96-well plate please organize samples vertically (ie. A1 through H1, A2 through H2 etc) and not horizontally (ie. A1 through A12, B1 through B12).
Please label samples in individual tubes or strips clearly by number (1-96).
How much sample do I need to submit?
| Assay | Minimum Volume |
|---|---|
| RNA | 2 uL |
| HS RNA | 3 uL |
| DNA 1000 | 2 uL |
| HS DNA 1000 | 3 uL |
| DNA 5000 | 2 uL |
| HS DNA 5000 | 3 uL |
| gDNA | 2 uL |
What do my TapeStation results mean?
The TapeStation result will really depend on what kind of sample you are testing. It will vary significantly if it is extracted RNA, an amplified DNA product, a fragmented library prepared for NGS, or genomic DNA. Below are some examples of common sample types and what the results often look like. If you're ever unsure about what your results mean, please contact us.
Amplicon Samples
For amplicons, you should notice a fairly pronounced peak at an approximate size of the amplicon that you are trying to amplify using PCR (714 in the above example). You will also notice a "Lower" and "Upper" peak that are added to each sample to provide a frame of reference for the TapeStation to determine peak sizes of your sample.
Fragmented libraries ex. for NGS
A fragmented library, such as those used in Illumina NGS applications will have a more gradual peak, along with peaks for the "Lower" and "Upper" markers. The peak is more gradual in this case as the DNA is fragmented into varying lengths. In this case the peak size (440 in the above example) is more indicative of an average size of the fragmented DNA.
RNA samples
You will notice that for RNA samples from eukaryotic organisms, there are three peaks: the first is the "Lower" marker, one labelled 18S and the other 28S. The 18S and 28S peaks correspond to RNA from the ribosomal subunits. In between these peaks, there are areas indicating the presence of RNA from non-ribosomal subunits.
RNA samples also have one more consideration: the RNA Integrity Number or RIN. In the above example, you can see there is a value of 9.7 which is highlighted in green. The RIN indicates how degraded a sample is and ranges from 1 to 10, with 10 being RNA that has very little degradation and 1 being a very degraded sample. This value is determined based upon a ratio of the 18S and 28S subunits.
Genomic DNA
Genomic DNA TapeStation will typically have two peaks, one indicating the "Lower" marker and a peak at a size approximately the size of the genome in your sample.