tagmentation washing buffer  (Boston BioProducts)

Journal: Genome Biology

Article Title: txci-ATAC-seq: a massive-scale single-cell technique to profile chromatin accessibility

doi: 10.1186/s13059-023-03150-1

Figure Lengend Snippet: txci-ATAC-seq generates high-quality single-cell ATAC libraries at high throughput. a Schematic of molecular details of txci-ATAC-seq library generation. b Experimental workflow for txci-ATAC-seq barnyard library generation. After 96-plex tagmentation, nuclei are overloaded on a 10X Chromium microfluidics device. Following nucleus encapsulation in the formed droplets, 10% of the GEMs can be used for quality control and the remaining 90% for data analysis. c-e) txci-ATAC-seq QC metrics for human (GM12878) and mouse (CH12) cell lines supplemented with SBS primer during in-droplet PCR. c “Knee” plot showing the unique reads (log 10 scale) against the rank of each barcode (log 10 scale) ordered from most unique reads (left) to least (right). The dashed line indicates the threshold (1000 reads) used to identify cell barcodes (orange points). d Scatter plots showing the number of unique reads mapped to either the human or mouse genome for both true and pseudo-barnyard experiments. Values were log 10 -transformed after adding a pseudo-count of 1 to all values. The percentage shown in the true barnyard panel (6.6%) represents the estimated collision rate. e Scatter plots showing the FRiDHS against the estimated complexity for each cell barcode detected as either mouse (blue) or human (red) cell. The estimated complexity is shown on a log 10 scale

Article Snippet: After quenching enzyme activity, the nuclei were pooled into a 12-tube strip and then transferred to a 15-ml conical tube preloaded with 400 μl tagmentation washing buffer (TMG, which contains 10 mM Tris acetate pH 7.8 (Boston BioProducts, Cat. BB-2412), 5 mM magnesium acetate (Sigma, Cat. 63,052-100ML), and 10% (v/v) glycerol (VWR, Cat. RC3290-32) diluted in nuclease-free water.

Techniques: High Throughput Screening Assay, Encapsulation, Control, Transformation Assay

Journal: Genome Biology

Article Title: txci-ATAC-seq: a massive-scale single-cell technique to profile chromatin accessibility

doi: 10.1186/s13059-023-03150-1

Figure Lengend Snippet: txci-ATAC-seq generates high-quality single-cell ATAC libraries at high throughput. a Schematic of molecular details of txci-ATAC-seq library generation. b Experimental workflow for txci-ATAC-seq barnyard library generation. After 96-plex tagmentation, nuclei are overloaded on a 10X Chromium microfluidics device. Following nucleus encapsulation in the formed droplets, 10% of the GEMs can be used for quality control and the remaining 90% for data analysis. c-e) txci-ATAC-seq QC metrics for human (GM12878) and mouse (CH12) cell lines supplemented with SBS primer during in-droplet PCR. c “Knee” plot showing the unique reads (log 10 scale) against the rank of each barcode (log 10 scale) ordered from most unique reads (left) to least (right). The dashed line indicates the threshold (1000 reads) used to identify cell barcodes (orange points). d Scatter plots showing the number of unique reads mapped to either the human or mouse genome for both true and pseudo-barnyard experiments. Values were log 10 -transformed after adding a pseudo-count of 1 to all values. The percentage shown in the true barnyard panel (6.6%) represents the estimated collision rate. e Scatter plots showing the FRiDHS against the estimated complexity for each cell barcode detected as either mouse (blue) or human (red) cell. The estimated complexity is shown on a log 10 scale

Article Snippet: Tagmented nuclei were then pooled into a single 15-ml conical tube; 5 ml of tagmentation wash buffer (TMG) was prepared consisting of a final concentration of 10 mM Tris acetate pH 7.5 (Sigma 93,352 and Sigma A6283, respectively), 5 mM magnesium acetate (Sigma M5661), and 10% (v/v) glycerol (Sigma G5516), diluted in PCR grade water; 1 ml of TMG was added on top of the chilled tagmented nuclei.

Techniques: High Throughput Screening Assay, Encapsulation, Transformation Assay