Specifications, Last Modified on October 5, 2022, Permalink
The Chromium™ Fixed RNA Profiling Solution produces Illumina® sequencer-ready libraries.
Please note that Dual Index Kit TS, Set A (PN-1000251) is required for use with the Fixed RNA Profiling Solution.
Supported Sequencers
Recommended Sequencing: Minimum 10,000 read pairs/cell*
Dual-Indexed Sequencing Run: Fixed RNA Profiling libraries are dual-indexed. We do not recommend sequencing these libraries with a single-index configuration.
PhiX Spike-In Recommendations: 5% for Multiplexed libraries; 1% for Singleplexed libraries
| Read | Read 1 | i7 Index | i5 Index | Read 2 |
|---|---|---|---|---|
| Purpose | Cell barcode & UMI | Sample Index | Sample Index | Insert |
| Length** | 28 | 10 | 10 | 90 *** |
*Adjust sequencing depth for the required performance or application. The Sequencing Saturation metric and curve in the Cell Ranger run summary can be used to optimize sequencing depth.
**Shorter transcript reads may lead to reduced transcriptome alignment rates. Cell barcode, UMI and Sample index reads must not be shorter than indicated. Any read can be longer than recommended. Additional bases in Sample index reads must be trimmed using cellranger mkfastq or Illumina's bcl2fastq prior to further analysis. Additional bases in Cell barcode or UMI reads will automatically be ignored by Cell Ranger.
*** The minimum required read length for Read 2 in Singleplex libraries is 50bp, while the minimum read length required for Read 2 in Multiplexed libraries is 76 bp.
Supported Sequencers
Recommended Sequencing: Minimum 5,000 read pairs/cell*
Dual Indexed Sequencing Run: Fixed RNA Profiling Feature Barcode libraries are dual-indexed. We do not recommend sequencing 10x Single Cell Fixed RNA Profiling Feature Barcode libraries with a single-index configuration.
We recommend pooling Fixed RNA Profiling Feature Barcode libraries with Fixed RNA Profiling Gene Expression libraries to maintain nucleotide diversity during sequencing.
| Read | Read 1 | i7 Index | i5 Index | Read 2 |
|---|---|---|---|---|
| Purpose | Cell barcode & UMI | Sample Index | Sample Index | Insert |
| Length** | 28 | 10 | 10 | 90 |
*Adjust sequencing depth for the required performance or application. The Sequencing Saturation metric and curve in the Cell Ranger run summary can be used to optimize sequencing depth for specific sample types.
**If sequencing Fixed RNA Profiling Cell Surface Protein libraries independently, they may also be sequenced in a 28 x 10 x 10 x 25 bp configuration.