CUTANA™ Nuclei Extraction Buffer

SKU: 21-1026
Pack Size: 100 Reactions


CUTANA Nuclei Extraction Buffer is the essential reagent for harvesting nuclei from cultured cells and tissues for use in CUT&RUN and CUT&Tag assays. This buffer is expected to be broadly compatible with eukaryotic cells and tissues. Below are examples of various cells and tissue types that have undergone successful nuclei isolation using this buffer:

mouse NIH3T3 fibroblast cells human A549 non-small cell lung cancer (NSCLC) cells
human K562 leukemia cells human NCI-H1299 non-small cell lung cancer (NSCLS cells)
human bone marrow derived macrophages human TIG-1 fetal lung cells
human monocyte derived macrophages human LoVo colorectal cancer cells
human MV-4-11 macrophage cells human LNCaP prostate carcinoma cells
human SUM149 triple negative breast cancer (TNBC) cells human renal primary cells
human GM24385 B-lymphocyte (aka HG002) cells human peripheral blood mononuclear cells (PBMCs)
human MCF7 breast cancer cells human intestinal tissue
human MDA-MB-231 breast cancer cells human HEPM embryonic cells
human SK-MEL-2 melanoma cells

The reagent is prepared by supplementing Pre-Nuclei Extraction Buffer with protease inhibitor and spermidine fresh on the day of use. Utilize this buffer in EpiCypher’s CUTANA Nuclei Extraction Protocol for CUT&RUN and CUT&Tag to obtain the highest quality nuclei for your genomic mapping assay.

Validation Data

Figure 1: Nuclei Extraction
Nuclei were extracted from 3 different cell types: K562, NIH3T3, and LNCaP using the CUTANA Nuclei Extraction Protocol for CUT&RUN and CUT&Tag. Top 3 panels show cells before extraction. Starting cells are viable (bright white and round). Bottom 3 panels show nuclei extracted using the CUTANA Nuclei Extraction Buffer as indicated by positive Trypan Blue staining.


Item Cat. No.
CUTANA Pre-Nuclei Extraction Buffer 21-1026a
1 M Spermidine 21-1026b

Required Materials Not Supplied

Protease inhibitor is not included. EpiCypher suggests using cOmplete , EDTA-free Protease Inhibitor Cocktail (Roche 11873580001).

Recommended Accessory Products

Item Cat. No.
CUTANA pAG-MNase for ChIC/CUT&RUN 15-1016/15-1116
CUTANA pAG-Tn5 for CUT&Tag 15-1017/15-1117

NOTE: CUTANA CUT&Tag Kit (14-1102/14-1103) already contains Nuclei Extraction Buffer

Technical Information

Store the Pre-Nuclei Extraction Buffer at 4ºC and the 1 M Spermidine at -20ºC. Components are stable for 6 months upon date of receipt.

Instructions for Use

Prepare Nuclei Extraction Buffer FRESH on day of use as outlined below. This master mix includes 20% extra volume to account for pipetting error; no additional buffer volume is needed.

Per CUT&RUN or CUT&Tag reaction, combine:

235 µL Pre-Nuclei Extraction Buffer (21-1026a)

0.13 µL 1M Spermidine (21-1026b)*

9.8 µL 25X Protease Inhibitor (Roche, prepare by dissolving 1 tablet in 2 mL water)**

*If preparing buffer for fewer than 8x reactions, dilute the 1M Spermidine stock 1:10 in molecular biology-grade water and add 1.3 µL per reaction.

**Leftover 25X Protease Inhibitor can be stored for 12 weeks at -20ºC

Follow instructions detailed in EpiCypher’s CUTANA Nuclei Extraction Protocol for CUT&RUN and CUT&Tag to isolate nuclei from suspension and adherent cells. The protocol includes key quality control checks to ensure nuclei are high quality before starting CUT&RUN or CUT&Tag.

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