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. 

We perform qPCR on all pools and 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. 

Pre-Sequencing QC Options

1. Pooling & 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
      • Unless stated otherwise, libraries are pooled equimolar
        • If pooling is not equimolar, provide the percentages of each sample or pool in the "Short Read Template"
    • Once pooled, the final pool will enter our Pre-Sequencing QC queue, and the steps in Option 2 will be completed before sequencing

 

2. QC on Pool

This option is for researchers submitting a library pool. 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 pool
    • 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. We perform qPCR as a service and guarantee to deliver at least 90% of Illumina's advertised optimal number of reads per lane/run. 
    • For faster processing, you can request to skip qPCR and our sequencing guarantee. 
      • Any failed runs or poor quality will have to be rerun as a new service. 

3. No QC

This option is for researchers that want to bypass qPCR prior to sequencing.

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.