View a description and download our example datasets below.

 

10k peripheral blood mononuclear cells (PBMCs) from 2 healthy donors with TotalSeq™-A Human Universal Cocktail

 

Cell Ranger 6.1.0 (10x Genomics)

 

Peripheral blood mononuclear cells (PBMCs) from 2 healthy donors were stained separately with TotalSeq™ hashtags and the TotalSeq™-A Human Universal Cocktail, V1.0 and then pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation, and sequencing were carried out according to our protocol: TotalSeq™-A Antibodies and Cell Hashing with 10x Single Cell 3' Reagent Kit v3.1 (Dual Index) Protocol.

  • 10,213 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

99dab79de03dc926d8e0c11926a45ab0

Feature Reference CSV

c9676eebc0f5b07a0ea9609c657f5d85

Output Files

 

Molecule info H5

4a87693f7daa3c3bf1c7e3d1536a0b7d

Feature bc matrix (filtered) Cell Ranger output fileMAS Compatible

8b0e50c273c4d051f26b715007e3da4a

Feature bc matrix (raw) Cell Ranger output file 

1ccf6b291b2b22a7f278def5cb50569d

Metrics summary

1f9cb40acd660b0c1891476658eaabfb

Web summary HTML

5c9f51a872c94a89e89af4670fc49661

 

 

10k peripheral blood mononuclear cells (PBMCs) from 2 healthy donors with TotalSeq™-B Human Universal Cocktail

 

Cell Ranger 6.1.0 (10x Genomics)

 

Peripheral blood mononuclear cells (PBMCs) from 2 healthy donors were stained separately with TotalSeq™ hashtags and the TotalSeq™-B Human Universal Cocktail, V1.0 and pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation and sequencing were carried out according to our protocol: TotalSeq™-B or -C with 10x Feature Barcoding Technology

  • 10,132 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

cf45624a4b10f691ed9b8395285c182f

Feature Reference CSV

04a9182ea5ef28a1f3e62b4785278e0c

Output Files

 

Molecule info H5

1eaa8c3b663b9d319a6d6bde7d158c87

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

23b3a0a1edb702c222210989f9103532

Feature bc matrix (raw) Cell Ranger output file       

967edf68e736e37781e91953a3f0e7ad

Metrics summary (link to file)

7a97aca87faeb450c2d140f556e303fe

Web summary HTML (link to file)

399d53bd3e086833f8c88e68bf4e45ba

 

 

12k peripheral blood mononuclear cells (PBMCs) from 2 healthy donors with TotalSeq™-C Human Universal Cocktail

 

Cell Ranger 6.0.2 (10x Genomics)

 

Peripheral blood mononuclear cells (PBMCs) from 2 healthy donors were stained separately with TotalSeq hashtags and the TotalSeq™-C Human Universal Cocktail, V1.0 and pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation and sequencing were carried out according to our protocol: TotalSeq™-B or -C with 10x Feature Barcoding Technology.

 

  • 12,062 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

80f8741fee05f474ee4c882f3a683896

Feature Reference CSV 

1a5243769226c527965bda6c1e7dacdf

Output Files

 

Molecule info H5

2ece3fcb7b91b288c55a4f6e25491a44

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

f753d67f907f252cc9b11903e5c8b165

Feature bc matrix (raw) Cell Ranger output file    

b935c50fccb3c04cdcd2f0b4c7de1bdb

Metrics summary

8ddf4dc0b2d648ceed573f13c1a610da

Web summary HTML

3e0f8dea471977c014c7e9e64c90deee

 

 

BEN-seq with 500k peripheral blood mononuclear cells (PBMCs) from 2 healthy donors with TotalSeq™- A Human Universal Cocktail

 

 

Peripheral blood mononuclear cells (PBMCs) from 2 healthy donors were activated with PMA and Ionomycin for 6 hours. For each donor, a total of two samples: the original resting cells, and the activated cells were collected. All four samples were stained with a 10% dilution of TotalSeq™-A Human Universal Cocktail, V1.0.

 

Staining, library preparation, and sequencing were carried out according to our protocol: Bulk Epitope and Nucleic Acid Sequencing (BEN-seq) Protocol

 

30 µL of the diluted libraries were sequenced to obtain paired end 30 bp (30x2) data on two MiniSeq High output 75 cycle kits (Illumina).

Data Files

md5sum

Activated cells FASTQs

2614f742084731b61ef32d2180a58891

Resting cells FASTQs

186f83b220ee77b66a9335cacb551a14

Feature reference CSV 

f3ecdb5568f5bb29b953d7d6c03a6c7f

 

File Naming Scheme

Each file is named using the following template: d10x_pbmc_all_act_d1r1_ADT_S11_R1_001.fastq.gz, where:

  • d10x refers to the samples being stained by 10x diluted TotalSeq™-A Human Universal Cocktail.
  • PBMC refers to the cell type (peripheral blood mononuclear cells, or PBMC).
  • all act refers to the sequenced sample contains 100% activated cells. This field is omitted for resting cells.
  • d1 refers to the cells come from Donor 1.
  • r1 refers to the sample being first of two replicates.
  • ADT refers to the library sequenced is from antibody-derived tags (ADT).

The remaining fields are identical to the fields used given by bcl2fastq to demultiplexed FASTQ data files.

 

 

 

16k splenocytes from 3 C57BL/6 mice with TotalSeq™-A Mouse Universal Cocktail

 

 

Cell Ranger 6.1.0 (10x Genomics)

 

Splenocytes harvested by mechanical dissociation from three C57BL/6 mice were stained separately with TotalSeq™ Hashtags and the TotalSeq™-A Mouse Universal Cocktail, V1.0 and pooled prior to GEM generation. Only resting cells in this analysis.

 

Staining, library preparation, and sequencing was carried out according to our protocol: TotalSeq™-A Antibodies and Cell Hashing with 10x Single Cell 3' Reagent Kit v3.1 (Dual Index) Protocol

 

  • 16,689 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

536ee8d52258f577e17ef2d72cf794bd

Feature Reference CSV

6b143d2372ae2e90e94d7d557ecbbfdf

Output Files

 

Molecule info H5

759c2607df676278c775a7cd30a29cf4

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

ee6c81bd52860d5f0dd8bd6c5194de88

Feature bc matrix (raw) Cell Ranger output file             

0d8ac948c9264bccfda50486e33d90a1

Metrics summary

a9b2a82cafbc7df61775bca38e666a9e

Web summary HTML

0650676c8e9b09ae00f27b27001a0c06

 

 

 

16k splenocytes from 3 C57BL/6 mice with TotalSeq™-B Mouse Universal Cocktail

 

Cell Ranger 6.1.0 (10x Genomics)

 

Splenocytes harvested by mechanical dissociation from three C57BL/6 mice were stained separately with TotalSeq™ hashtags and the TotalSeq™-B Mouse Universal Cocktail, V1.0 then pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation, and sequencing were carried out according to our protocol: TotalSeq™-B or -C with 10x Feature Barcoding Technology.

 

  • 18,290 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp i7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

73954ce271142a27259a96751de1d1e8

Feature Reference CSV

86561c17adcf75ad3e12f679d29f8640

Output Files

 

Molecule info H5

5e1756373d68efd50fabb2be856cef6e

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

e1273f56463ceb9df95a7a00fd01de85

Feature bc matrix (raw) Cell Ranger output file              

a6341fcd93c5073aa35f6631ac01123e

Metrics summary

4cb2e7164d6454011dabf351bb28c3ef

Web summary HTML

5f1ef57aaf432c0103eff3cf23e6b16e

 

 

 

16k splenocytes from 3 C57BL/6 mice with TotalSeq™-C Mouse Universal Cocktail

 

 

Cell Ranger 6.1.0 (10x Genomics)

 

Splenocytes harvested by mechanical dissociation from three C57BL/6 mice were stained separately with TotalSeq™ Hashtags and the TotalSeq™-C Mouse Universal Cocktail, V1.0 and then pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation, and sequencing were carried out according to our protocol: TotalSeq™-B or -C with 10x Feature Barcoding Technology.

 

  • 13,566 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO). 
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp i7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

fc56599725581d1f0e3b3ddefa56c617

Feature Reference CSV

e2eadbe12f1a03e584b6ba997aaf5f26

Output Files

 

Molecule info H5

fad175b3288816a9d75fd68c02a7d282

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

50de00dc86050c41caab0afded8d6b68

Feature bc matrix (raw) Cell Ranger output file              

ac95a42da0334eb89e3110572a1e4075

Metrics summary

3db8b127408ffe409cdaa9d4d434a9a6

Web summary HTML

1d2599859367f469356f6aeb347e93db

 

 

 

 

12k Splenocytes from 2 C57BL/6 mice with TotalSeq™-B Mouse Myeloid Cocktail

 

 

Cell Ranger 7.1.0 (10x Genomics)

 

Splenocytes were harvested by enzymatic dissociation of spleens from two C57BL/6 mice. T and B cell populations were depleted using MojoSort™ anti-PE Nanobeads, PE anti-mouse CD90.2 (Thy-1.2), and PE anti-mouse CD19. Cells from each mouse were stained separately with TotalSeq™ Hashtags, the TotalSeq™-B Mouse Myeloid Cocktail, V1.0, and then pooled prior to GEM generation. Only resting cells were included in this analysis.

 

Staining, library preparation, and sequencing was carried out according to our protocol: TotalSeq™-B or -C with 10x Feature Barcoding Technology.

 

  • 10,022 cells detected
  • Sequenced on Illumina NovaSeq 6000 with approximately 36,000 reads per cell (25,000 RNA, 10,000 ADT, 1,000 HTO).
  • 28bp read 1 (16bp Chromium barcode and 12bp UMI), 91bp read2 (transcript), and 8bp I7 sample barcode
  • run with --expect-cells= 10,000

 

Input Files

md5sum

FASTQs

e15ced63c4488015ad739fe1afaa0953

Feature Reference CSV

1a1fcdbd82597e5d54bf6fdb3e6f5895

Output Files

 

Molecule info H5

620c00a8cd75b023ddf8b2e059197b98

Feature bc matrix (filtered) Cell Ranger output file, MAS Compatible

5df44f651a0ca6390553aa35b459b4fb

Feature bc matrix (raw) Cell Ranger output file              

1eeff2004c277d0796f7f4e15ccdcccc

Metrics summary

8e9d783782e73def44bdcd91f48a94af

Web summary HTML

ae2186911089addaf0dfa6ca9cfbf1e3

 

 

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