title: Adult1_S32
organism: Macaca fascicularis
description: Protocols: Macaca fascicularis tetis tissues were incubated in dissociation buffer containing 200 U/mL collagenase I (Gibco, 17100017), for up to 5 mins with gentle agitation to dissociate tubules. The supernatant, containing mostly interstitial cells, was removed and fresh dissociation buffer containing collagenase, with 2.4 U/mL dispase II (Gibco, 17105041) and 1 mM CaCl2 in Hanks Balanced Salt Solution (HBSS) was added to the sample, and incubation was performed at 37 °C for 30 minutes in a water bath. Tubes were agitated with shaking every 5 minutes until tissue is dissociated fully into single cells. Enzymatic digest was quenched by the addition of 10 % FBS. Cells are then washed and resuspended in PBS, and filtered through a 40 μM strainer in order to obtain a suspension of single-cells. 4.8 ul of single-cell suspension at a concentration of ~1,000,000 cells/ml was loaded into one channel of the Chromium Single Cell Chip B (10X Genomics®), aiming for a recovery of 3000-4000 cells. Chromium Single Cell 3' Library & Gel Bead Kit using version 3 chemistry (10X Genomics, PN-1000092) was used for single cell barcoding, cDNA synthesis and library preparation, following manufacturer's protocol (Chromium Single Cell 3ʹ Reagent Kits v3 User Guide, Revision B).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2020-08-13T04:07:19Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2020-04-23T17:13:30Z
attributes.tag: External Id
attributes.value: SAMEA6799513
attributes.tag: INSDC center name
attributes.value: KK Women's and Children's Hospital, Singapore
attributes.tag: INSDC first public
attributes.value: 2020-08-13T04:07:19Z
attributes.tag: INSDC last update
attributes.value: 2020-04-23T17:13:30Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-8979:Adult1_S32
attributes.tag: age
attributes.value: 48
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: animal reproductive system cell
attributes.tag: common name
attributes.value: crab-eating macaque
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: immunophenotype
attributes.value: unsorted
attributes.tag: individual
attributes.value: Adult1
attributes.tag: organism part
attributes.value: testis
attributes.tag: sample name
attributes.value: E-MTAB-8979:Adult1_S32
attributes.tag: scientific_name
attributes.value: Macaca fascicularis
attributes.tag: sex
attributes.value: male