10x
Support
Search Support
Contact us
10x Genomics Support/Single Cell Gene Expression/Documentation/Sequencing/

Sequencing Requirements for Single Cell 3' HT

Single Cell 3' HT v3.1 Dual Index Gene Expression

The Chromium™ Single Cell Gene Expression HT Solution with Feature Barcode technology produces Illumina® sequencer-ready libraries.

Supported Sequencers

  • Illumina® NovaSeq
  • Illumina® HiSeq 3000/4000
  • Illumina® HiSeq 2500 Rapid Run
  • Illumina® NextSeq 500/550
  • Illumina® NextSeq 1000/2000
  • Illumina® MiSeq

Recommended Sequencing: Minimum 20,000 read pairs/cell*

Dual Indexed Sequencing Run: Single Cell 3' HT v3.1 Dual Index libraries are dual-indexed. We do not recommend sequencing 10x Single Cell 3’ HT v3.1 Dual Index libraries with a single-index configuration. See: Can single index and dual index libraries be pooled for sequencing?

ReadRead 1i7 Indexi5 IndexRead 2
PurposeCell barcode & UMISample IndexSample IndexInsert
Length**28101090

*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 (note: this metric was named cDNA PCR Duplication in Cell Ranger 1.1 or earlier).

**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 1.3 or later.

Single Cell 3' HT v3.1 Dual Index Gene Expression with Feature Barcode technology

Supported Sequencers

  • Illumina® NovaSeq
  • Illumina® HiSeq 3000/4000
  • Illumina® HiSeq 2500 Rapid Run
  • Illumina® NextSeq 500/550
  • Illumina® NextSeq 1000/2000
  • Illumina® MiSeq

Recommended Sequencing: Minimum 5,000 read pairs/cell*

Dual Indexed Sequencing Run: Single Cell 3' HT v3.1 Dual Index Feature Barcode libraries are dual-indexed. We do not recommend sequencing 10x Single Cell 3’ HT v3.1 Dual Index Feature Barcode libraries with a single-index configuration.

We recommend pooling Single Cell 3' HT v3.1 Dual Index Feature Barcode libraries with Single Cell 3' HT v3.1 Dual Index Gene Expression libraries to maintain nucleotide diversity during sequencing.

ReadRead 1i7 Indexi5 IndexRead 2
PurposeCell barcode & UMISample IndexSample IndexInsert
Length**28101090

*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 (note: this metric was named cDNA PCR Duplication in Cell Ranger 1.1 or earlier).

**If sequencing 3' HT v3.1 Dual Index Cell Surface Protein libraries independently, they may also be sequenced in a 28 x 10 x 10 x 25 bp configuration. We do not recommend sequencing Single Cell 3’ HT v3.1 Dual Index CRISPR Screening or Cell Multiplexing Libraries independently.

Document Type
Specifications

Last Modified
August 9, 2021