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Item

10x PN

Preparation & Handling

Storage

Equilibrate to Room Temperature

ATAC Buffer C

2001581

Thaw, vortex, centrifuge briefly.

−20°C

20X Nuclei Buffer

Concentrated 20X stock; dilute 1:20 in nuclease-free water before use. (See Prepare Diluted Nuclei Buffer)

2000207

Thaw. Vortex, centrifuge briefly.

−20°C

Reducing Agent B

2000087

Thaw, vortex, verify no precipitate, centrifuge briefly.

−20°C

Nuclease-free Water

Place on Ice

RNase Inhibitor

2001488

−20°C

For Post Transposition Wash Buffer, use ONLY PN-2001488

For Diluted Nuclei Buffer, use PN-2001488 or equivalent

ATAC Enzyme B

2001582/

2001583

Centrifuge briefly, pipette mix

−20°C

Nuclei*

in Diluted Nuclei Buffer

(See below to Prepare Diluted Nuclei Buffer)

*Refer to protocols for Nuclei Isolation for ATAC + Gene Expression (CG000365, CG000375, CG000505). Adhering to these protocols is critical for optimal assay performance.

Prepare Diluted Nuclei Buffer Maintain at 4°C

The use of the Tris-based Diluted Nuclei Buffer for nuclei suspension is critical for optimal assay performance. The composition of the Diluted Nuclei Buffer, including Magnesium concentration, has been optimized for the Transposition and Barcoding steps. Suspension of nuclei in a different buffer may not be compatible with the downstream protocol steps.

Total	-	-	1000 µl
	Stock	Final	Volume (µl)
20X Nuclei Buffer (PN-2000207; equilibrated to room temp.)	20X	1X	50 μl
Reducing Agent B (PN-2000087; equilibrated to room temp.)	1,000 mM	1 mM	1 µl
RNase Inhibitor (Use PN-2001488 or equivalent. Confirm vendor-specific stock concentration)	40 U/µl	1 U/µl	25 µl
Nuclease-free Water	-	-	924 µl