Required reagents included in kit

Kit components are stable for 6 months upon date of receipt. Store as outlined below.

Store at room temperature (RT) upon receipt:

Item Catalog Number Notes
8-strip Tubes 10-0009a Enables use of multi-channel pipettors.
DNA Cleanup Columns *for kit Versions 1-3 10-0010 Use with the DNA Collection Tubes.
DNA Collection Tubes *for kit Versions 1-3 10-0011 Use with the DNA Cleanup Columns.
DNA Binding Buffer *for kit Versions 1-3 21-1008 Before first use, add 6.9 mL isopropanol. WARNING: Contains toxic ingredients (see Safety Data Sheet).
DNA Wash Buffer *for kit Versions 1-3 21-1009 Before first use, add 20 mL of ≥95% ethanol.
DNA Elution Buffer *for kit Versions 1-3 21-1010 Use to elute final CUT&RUN DNA.
SPRIselect Reagent Manufactured by Beckman Coulter **for kit Version 4 21-1405 DO NOT FREEZE. Reagent is slightly viscous. Thoroughly mix prior to use and pipette carefully to ensure correct volume is transferred. Used to purify CUT&RUN-enriched DNA from assay supernatant.
0.1X TE Buffer **for kit Version 4 21-1025 Used to elute CUT&RUN-enriched DNA.
0.5 M EDTA 21-1006 250X concentration. Use to prepare Antibody Buffer FRESH for each experiment.
100 mM Calcium Chloride 21-1007 Activates chromatin-tethered pAG-MNase to cleave DNA.

Store at 4°C upon receipt:

Item Catalog Number Notes
ConA Beads 21-1401 DO NOT FREEZE. Concanavalin A (ConA) beads are used for immobilizing nuclei or cells. Because ConA can cause immune cell activation, it is recommended to use nuclei for immune cell studies (see this article).
E. coli spike-in DNA 18-1401 100 ng lyophilized E. coli DNA. Before first use, quick spin and reconstitute in 200 μL DNase-free water (0.5 ng/μL). Add to reactions for sequencing normalization. NOTE: After reconstitution, store at -20°C.
Bead Activation Buffer 21-1001 Use to prepare ConA beads prior to sample immobilization.
Pre-Wash Buffer 21-1002 Use to prepare Wash, Cell Permeabilization, and Antibody Buffers FRESH for each experiment.
Stop Buffer 21-1003 3X concentration. Use to terminate pAG-MNase cleavage activity.

Store at -20°C upon receipt:

Item Catalog Number Notes
5% Digitonin 21-1004k Thaw at RT. Use to prepare Cell Permeabilization and Antibody Buffers FRESH for each experiment. Final Digitonin concentration should be optimized for each sample type, see this article.
1 M Spermidine 21-1005 2,000X concentration. Use to prepare Wash Buffer FRESH for each experiment.
SNAP-CUTANA™ K-MetStat Panel 19-1002k SMALL VOLUME: quick spin before use. Pipette to resuspend - DO NOT VORTEX. Panel of biotinylated nucleosomes coupled to streptavidin-coated magnetic beads. Pair with IgG and H3K4me3 control antibodies. Sufficient for 20 reactions. See this section for more information.
Rabbit IgG Negative Control Antibody 13-0041k SMALL VOLUME: quick spin before use. 0.5 mg/mL stock. Add 1μL to negative control reactions. Sufficient volume for 10 reactions.
H3K4me3 Positive Control Antibody 13-0042k SMALL VOLUME: quick spin before use. 0.5 mg/mL rabbit mixed monoclonal antibody. Add 1 μL to positive control reactions. Sufficient volume for 10 reactions.
pAG-MNase 15-1016 20X concentration. Proteins A and G (pAG) bind antibodies of various isotypes and host species including total IgG for rabbit, mouse, goat, donkey, rat, guinea pig, horse, and cow.

SPRIselect Reagent

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