Sample Type | Optimal | Range (total RNA) |
---|---|---|
Cell fractions | N/A | 500 – 50,000 cells |
Tissue (fresh/frozen) | N/A | up to 1 mg |
Blood Microsamples | N/A | 30 ul |
As shown in the Table above, for a small number of cells, such as immune cell fractions isolated by FACS or magnetic beads (e.g. 5,000-50,000 cells), and tissue samples (up to ~1 mg), we recommend using cells directly without RNA purification step. Using sorted cells/tissues increases the sensitivity of CDR clonotype detection as compared to RNA-isolated samples. Refer to the guidelines in this section for the preparation of these samples.
To use small samples directly in the assay, follow the procedure below:
- Purify immune cells or prepare dissociated tissue samples (e.g., by collagenase treatment). Adjust the volume of each sample to a total of 14 ul. Depending on your cell collection method, this may require spinning down the cells and removing excess supernatant over 14 ul.
- When you are ready to start the DriverMap AIR Assay, prepare the Hybridization Buffer Master Mix as described in Step 1 of the Hybridization Procedure, add N-Lauroyl Sarcosine to 0.3% (0.7 µl of 20% N-Lauroyl Sarcosine), and then set up the Hybridization reaction with your sample as described.
- Follow the standard protocol for hybridization, AMP purification, cDNA synthesis, and amplification steps.
Last modified:
20 July 2023
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