The Kit protocol consists of a 2-step PCR process to first amplify the CDR3 of a rearranged human TCRB locus and then to add Illumina®-specific adapter sequences to the TCRB locus amplicon. Each PCR step also includes a bead-based cleanup. We recommend quality control steps after both PCR steps. After the second PCR quality control step, you'll pool your samples, quantify the sequencing library present in the pool, and then load your samples on an Illumina MiSeq® or NextSeq® instrument for sequencing.

Using our data transfer utility (Macintosh®, Windows®, or Linux®) your raw sequence data is transferred to our servers. Once processed, your data will be available in your immunoSEQ® Analyzer workspace. You can then analyze, visualize, and share your data.

Download the immunoSEQ Kit Workflow.

Each Kit component is labeled with an expiration date. If properly stored and used in accordance with the instructions set forth in the Kit Manual, immunoSEQ® Kit reagents are covered by a limited warranty through the earliest expiration date marked on its components. The QIAGEN® Multiplex PCR Plus Kit is subject to separate warranties that might be offered by QIAGEN.

The number of samples that can be pooled in one sequencing run is dependent on the sample type and the sequencing machine being used. Roughly, a 96-well Kit will need to be batched into 3-5 MiSeq® runs. Whereas the entire 96-well plate could potentially be run on a single NextSeq® run. These approximations are based on the assumption that optimal cluster density was achieved for the sequencing run.

Additional information on sample pooling can be found in the immunoSEQ Kit Manual (download requires Analyzer login).

Log into your immunoSEQ Analyzer account and locate your Kit order on the Order Dashboard in the upper left of your screen. Click on the "Create Manifest" link next to the Kit order you are using. You can also access the manifest through the “Orders” link in the main menu (located in the upper left of the screen). From the orders page, click on the '+' sign to expand the order and then edit manifests for the kits. Make sure that the UID number next to the “Create Manifest” button corresponds to the UID on the bottom of your kit. The button will read “Edit Manifest” if you have worked with that Kit in the past.

Starting on the PCR PLATE tab, enter new samples one at a time by clicking the NEW SAMPLE button. Then assign each sample to a specific position of the PCR Plate. You can assign one sample to multiple wells to indicate the individual reactions for the given sample. Please refer to our Sample Naming Criteria when naming your samples.

The next step is to specify each sequencing batch in the POOLS tab. Create a new pool using the NEW POOL button, click on all relevant wells to select specific samples, and then click on "Assign to Pools."

Lastly, click on the SAMPLE SHEETS tab to download the sample sheet for your specific pool. Depending on the sequencing platform being used (MiSeq® versus NextSeq®), the sample sheet is necessary for optimal sequencing and data transfer. Please refer to you the Kit manual for specific instructions.

Each Kit has a unique ID (UID) that can be found on the bottom of the Kit box or by looking at your Kit order in the Analyzer.