title: GSM4659151: HIVandART1; Homo sapiens; RNA-Seq
library_construction_protocol: Frozen mature monocytes were thawed by placing in a 37o C water bath without agitation until only a small ice crystal was left in the vial. One ml of pre-warmed DNase supplemented media (RPMI supplemented with 20% FBS and 1% DNase I (Sigma, Cat# 10104159001)) was then added using filtered tips. The samples were gently mixed by pipetting and allowed to rest for 5 minutes at room temperature. After the 5 minutes, the contents of each vial were transferred to 10mL of pre-warmed DNase supplemented media and incubated for an additional 5 minutes at room temperature. Cells were then centrifuged, supernatant was aspirated, and the mature monocytes were resuspended in 1 mL of RPMI supplemented with 20% FBS and incubated for 15 minutes at 37o C with 5% CO2. After incubation cells were counted, centrifuged, and divided into two subsets, one used for staining post-freeze/thawing to verify viability on the day of scRNAseq and the other for scRNAseq. The scRNAseq cDNA libraries were prepared following the user guide manual provided by the 10X Genomics and Chromium Single Cell 3' Reagent Kits (v2 for HIV-infected ART-treated mature monocytes and v3 for HIV-infected mature monocytes). The libraries for the three HIV-infected ART-treated independent biological samples were run each individually on four lanes of sequencing on an Illumina Nextseq 550 instrument. For the HIV-infected independent biological samples, they were each divided in half for library preparation. The pooled libraries for one of the two HIV-infected independent biological samples were run on four lanes for sequencing on an Illumina Nextseq 550, and the pooled libraries for the second HIV-infected independent biological sample were run on two lanes on an Illumina Novaseq instrument. For the uninfected independent biological samples, they were each divided in half for library preparation. The pooled libraries were run on two lanes for sequencing on an Illumina Novaseq instrument.
attributes.tag: GEO Accession
attributes.value: GSM4659151