CUTANA™ E. coli Spike-in DNA

Catalogue Number: 18-1401-EPC

Manufacturer:	EpiCypher
Physical state:	Lyophilized
Type:	Other DNA and RNA
Shipping Condition:	Dry Ice
Storage Condition:	2-8°C
Unit(s):	100 ng

Overview
Datasheets

Description

Description: Fragmented DNA derived from Escherichia coli (E. coli) can be used as a spike-in control for experimental normalization in Cleavage Under Targets and Release Using Nuclease (CUT&RUN). CUT&RUN normalization was originally performed with carryover E. coli DNA from MNase preparation [1], however pure preps of enzyme require post hoc DNA spike-in. CUTANA™ E. coli Spike-in DNA contains sufficient material for 100-200 CUT&RUN reactions (range for high and low abundance targets).

Additional Text

Application Notes

Prior to opening, pellet DNA by quickly spinning in a benchtop microfuge. Reconstitute in 200 µL DNase free water (0.5 ng/µL). Vortex tube on all sides to ensure complete resuspension. Quick spin in benchtop microfuge prior to use. Use in CUT&RUN: See full protocol: www.epicypher.com/protocols 1. Use with CUTANA™ pAG-MNase for ChIC/CUT&RUN (EpiCypher 15-1016), which has very low E. coli DNA carryover. 2. Add 1-2 µL* (0.5-1 ng) E. coli Spike-in DNA directly to the Stop Buffer, which quenches calcium-mediated pAG-MNase DNA digestion. Gently vortex to mix, and add to samples. *NOTE: Based on target abundance and antibody efficiency, the amount of E. coli DNA may need to be adjusted. Aim for spike-in DNA to comprise ~1% (0.2-5%) of total sequencing reads (Figure 2). 3. After sequencing, align reads to the reference genome of experimental samples (e.g. human) as well as to E. coli genome. 4. Normalize data by following recommendations in the CUT&RUN Protocol: www.epicypher.com/protocols.

Storage Note

Stable for 2 years at 4°C from date of receipt. After resuspending, aliquots should be stored at -20°C.

18-1401 Datasheet

Price

£140.00

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