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Pre-Sequencing QC

The Bauer Core offers Pre-Sequencing QC to ensure proper loading on our Illumina sequences. Pre-sequencing QC includes TapeStation and qPCR. 
 
When we perform qPCR as a service, we guarantee to deliver at least 90% of Illumina's advertised optimal number of reads per lane/run as long as the sample passes our QC metrics (concentration >1nM and expected library peak size). 
 
**Note:If you want to guarantee pooling distributions, you must request a MiSeq QC run in addition to your flow cell or lane. 
 

Below are options for submitting samples for sequencing and Pre-Sequencing QC. Determine which option is best for your project and submit the corresponding requests in minilims. 

If you are unsure which option fits your needs, please contact Core Staff.

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Pre-Sequencing QC Options

Option 3 | No Pre-Sequencing QC

This option is for researchers submitting quantification data (either nM or ng/ul & average bp) for their pool. If you need the core to quantify your pool, see Option 2. 

While this option is the quickest turnaround time for sequencing, it does not come with any output guarantees. The researcher provides the final pool concentration used for loading the sequencer in their sequencing submission. Therefore, should your run fail, we require another sequencing request to process the rerun.

Option 2 | Pre-sequencing QC (TS & qPCR)

This option is for researchers submitting a library pool and would like the core to quantify prior to sequencing. Pre-Sequencing QC includes TapeStation and qPCR of each pool. If you need the core to pool your libraries before Pre-Sequencing QC, see Option 1.

  • TapeStation: determine the average fragment size of your libraries
    • Tapestation data verifies successful library construction and determines the average fragment size. The average fragment size is required to calculate the molar concentration after qPCR. 
  • qPCR: accurately quantify sequencing libraries
    • This is the most accurate sample quantification method since only adapter-ligated inserts will be counted. When we perform qPCR as a service, we guarantee to deliver at least 90% of Illumina's advertised optimal number of reads per lane/run. 

Option 1 | Library QC: Pooling & Pre-Sequencing QC

This option is for researchers submitting individual samples that need pooling prior to Pre-Sequencing QC. If your samples are already pooled, see Option 2. If you do not need pooling or Pre-Sequencing QC, see Option 3. 

  • Pooling: Pool libraries equimolar from TapeStation data
    • Tapestation data is used to determine the concentration of final libraries before pooling
      • Submit a TapeStation service request quantity will be number of submitted libraries for pooling
      • Unless stated otherwise, libraries are pooled equimolar
        • If pooling is not equimolar, provide the percentages of each sample or pool
    • Once pooled, the final pool will enter our Pre-Sequencing QC queue, and the steps in Option 2 will be completed before sequencing
See our fees page under "Quality Control & Reagents" for details.