# Cell input and volume loading recommendations for Illumina Single Cell 3’ RNA Prep

**Cell and Nuclei Loading Parameters**

Cell and nuclei loading in Illumina Single Cell 3’ RNA Prep kits is optimized to balance high recovery rates and data quality. The table below summarizes optimal loading parameters by kit size.

| Kit Size | Optimal Cell or Nuclei Input\* | Expected Capture Rate | Cell Loading Volume (µl) | RNase Inhibitor Volume (µl) | <p>Target<br>Concentration</p> |
| -------- | ------------------------------ | --------------------- | ------------------------ | --------------------------- | ------------------------------ |
| **T2**   | 5,000                          | ≥ 2,000               | 4                        | 1                           | 1,250 live cells/µl            |
| **T10**  | 17,000                         | ≥ 10,000              | 5                        | 1                           | 3,400 live cells/µl            |
| **T20**  | 40,000                         | ≥ 20,000              | 8                        | 1 - 2                       | 5,000 live cells/µl            |
| **T100** | 200,000                        | ≥ 100,000             | 20                       | 4                           | 10,000 live cells/µl           |

*\*Exceeding the recommended input increases the risk of a multiplet rate above 8%.*

**Notes:**

* Always include RNase inhibitor in the total loading volume.
* Refer to reference guides for loading volumes to ensure optimal capture efficiency.

**Cell and Nuclei Loading Volume** **Considerations**

* Adjust the target concentration if increasing or decreasing input.
* The total loading volume, including RNase inhibitor, must not exceed the specified limit for each kit size. Exceeding recommended volumes may reduce capture efficiency.
* If cells or nuclei cannot be concentrated to the target level, a lower concentration is generally acceptable as long as the total volume remains within recommended limits.

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| *For any feedback or questions regarding this article (Illumina Knowledge Article #9473), contact Illumina Technical Support* [*techsupport@illumina.com*](mailto:techsupport@illumina.com?subject=Question%2FFeedback%20Regarding%20Illumina%20Knowledge%20Article%20#000009473%20-%20Library%20Preparation%20\&body=Dear%20Illumina%20Technical%20Support,%0D%0A%0D%0A)*.* |


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