#  Pre-Sequencing QC 

 



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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.**

**\*\*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](https://bauer-minilims.rc.fas.harvard.edu/minilims).

If you are unsure which option fits your needs, please contact [Core Staff](mailto:illumina_submission-list@lists.fas.harvard.edu).



 

##  Pre-Sequencing QC Options 

 



 1. Pooling & QC 2. QC on Pool 3. No QC 

## 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 
        - 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](/fees) under "Quality Control & Reagents" for details. [Submit TapeStation/Library Pooling and Pre-Sequencing QC Requests](https://bauer-minilims.rc.fas.harvard.edu/minilims/plugins/Core/make_form.php?type=Reagent_Request&name=new)

 



 

 

 

## 2. QC on Pool

 

 

 *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**.

 [Submit a Pre-Sequencing QC Request](https://bauer-minilims.rc.fas.harvard.edu/minilims/plugins/Core/make_form.php?type=Reagent_Request&name=new)

 



 

 

 

## 3. No 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. [Submit a Sequencing Submission](https://bauer-minilims.rc.fas.harvard.edu/minilims//plugins/Core/submit_batch.php?parenttype=Submission&childtype=Sample&name=new)