PIXUL™ Chromatin Input Prep Kit Manual
Catalog No. 53134(version A1)

Active Motif North America
1914 Palomar Oaks Way, Suite 150
Carlsbad, California 92008, USAToll free:877 222 9543
Telephone:
760 431 1263
Fax:
760 431 1351
Active Motif Europe
Waterloo Atrium
Drève Richelle 167 – boîte 4
BE-1410 Waterloo, Belgium
UK Free Phone:		
0800 169 31 47
France Free Phone:
0800 90 99 79
Germany Free Phone: 0800 181 99 10
Telephone:		+32 (0)2 653 0001
Fax:		
+32 (0)2 653 0050
Active Motif Japan
Azuma Bldg, 7th Floor
2-21 Ageba-Cho, Shinjuku-KuTokyo, 162-0824, Japan
Telephone:+81 3 5225 3638
Fax:
+81 3 5261 8733
Active Motif China
787 Kangqiao Road
Building 10, Suite 202, Pudong District
Shanghai, 201315, China
Telephone:(86)-21-20926090
Hotline:400-018-8123
Copyright 2020 Active Motif, Inc.
www.activemotif.com

TABLE OF CONTENTS

Page

Overview . .  .  .  .  .  . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Kit Components and Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Reagents for Chromatin Input Prep  . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Additional Materials Required . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Protocol . .  .  .  .  .  .  . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Technical Services  . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

Information in this manual is subject to change without notice and does not constitute a commitment on thepart of Active Motif, Inc. It is supplied on an “as is” basis without any warranty of any kind, either explicit orimplied. Information may be changed or updated in this manual at any time.
This documentation may not be copied, transferred, reproduced, disclosed, or duplicated, in whole or in part,without the prior written consent of Active Motif, Inc. This documentation is proprietary information andprotected by the copyright laws of the United States and international treaties.
PIXUL™ is sold under an exclusive license to patents owned by Matchstick Technologies Inc. and University ofWashington, specifically Matula, T.J.; Bomsztyk, K.; Darlington, G.P., Maxwell, A.D.; MacConaghy, B.E., Reed, J.
“Ultrasound system for shearing cellular material”. US Patent US10809166. European Patent EP3169451.
The manufacturer of this documentation is Active Motif, Inc.
© 2020 Active Motif, Inc., 1914 Palomar Oaks Way, Suite 150; Carlsbad, CA 92008. All rights reserved.
All trademarks, trade names, service marks or logos referenced herein belong to their respective companies.
For Research Use Only. Not for use in diagnostic procedures.

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Overview
This kit is designed to analyze yield and sonication efficiency of the chromatin sheared with thePIXUL™ Chromatin Shearing Kit (Cat. No. 53132) using the PIXUL™ Multi-Sample Sonicator (Cat. No.
53130).
The kit contains sufficient reagents to prepare ChIP-ready chromatin for 96 IP reactions.

PIXUL Advantages• Process 1 - 96 samples in parallel• Extremely consistent sonication• Up to 12 different sonication conditions per run
product
PIXUL™ Chromatin Input Prep Kit

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1formatcatalog no.

1 X 96 reactions

53134

Kit Components and Storage
The kit contains sufficient reagents to prepare analyze ChIP-ready chromatin for 96 IP reactions.
The reagents in this kit have multiple storage temperatures. Please store components according tothe storage conditions below. All reagents are guaranteed stable for 6 months from date of receipt
when stored properly.
Reagents

QuantityStorage

SPRI Beads

3.98 mL4°C

Elution Buffer AM4

1.1 mLRT

Proteinase K

56 µL

-20°C

RNase A

2 X 45 µL-20°C

Low EDTA TE

2 X 1.2 mL

RT

Additional Materials Required• 100% Ethanol• Nuclease-free water• 1X PBS• Microcentrifuge tubes• PCR tube strips (300 µL well volume capacity)• Centrifuge to spin 96-well plate• Thermal cycler• 96-well plate magnet or bar magnet• Single channel and multi-channel pipettors and tips
• Hemocytometer and light microscope

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2

PIXUL Chromatin Input Prep Kit Protocol
Buffer Preparation1.

Prepare TE-RNase by combining 95 µL Low EDTA TE and 5 µL RNase A (10µg/µL).

2.

Prepare Elution Buffer with Proteinase K (EB-PK) by combining 95 µL Elution Buffer AM4 with 5µL Proteinase K (10µg/µL).

3.

Prepare 80% ethanol (4.8 mL 100% ethanol + 1.2 mL nuclease-free water).

Reverse Crosslinking
1.

Remove 10 μL of the chromatin that has been sonicated with the PIXUL Chromatin Shearing Kit(cat. no. 53132) and transfer to a PCR tube, strip-well, or 96-well plate2.

Add 10 µL TE-RNase to each sample and incubate for 30 minutes at 37°C.

3.

Add 10 µL EB-PK to each sample and incubate for 30 minutes at 55°C, followed by 90 minutes
at 65°C.

DNA Purification using SPRI Beads
Take SPRI Beads out of 4°C and warm to room temperature by simply placing the vial of SPRI beads
on the benchtop.
1.

Add 0.5 µL RNase A to each well and incubate for 30 minutes at 37°C.

2.

Add 5 µL Proteinase K to each well and incubate for 30 minutes at 55°C.

3.

Add 120 µL (1.2 volumes) of SPRI Beads to each well. Pipette the bead-lysate mixture 10 times
to mix, and incubate for 5 minute at room temperature.

4.

Place plate on a magnet for 5 minutes to collect beads, and then aspirate supernatant.

5.

Keeping the plate on the magnet, add 180 µL of 80% ethanol and incubate for 30 seconds atroom temperature.

6.

Aspirate the supernatant and perform a second wash with 180 µL of 80% ethanol.

7.

Completely aspirate the supernatant and air dry the beads until they are no longer shiny (2-5minutes).

8.

Remove the plate from the magnet and elute DNA by adding 50 µL of Low EDTA TE.

9.

Pipette the beads 10 times and incubate for 3 minutes at room temperature.

10. Place tube or plate back on the magnet for 5 minutes to collect beads.
11.

Carefully remove eluted DNA in the supernatant and transfer to a new tube or plate.

12.

Analyze gDNA yield using a spectrophotometric (Nanodrop) or fluorometric (Qubit) methodof your choice and analyze fragmentation efficiency using either agarose gel electrophoresis,
Agilent TapeStation, or Agilent Fragment Analyzer. For most applications, we recommend
shearing DNA to 200 - 600 bp.
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3Technical ServicesIf you need assistance at any time, please call Active Motif Technical Service at one of thenumbers listed below.
Active Motif North America
1914 Palomar Oaks Way, Suite 150
Carlsbad, CA 92008, USA
E-mail: tech_service@activemotif.com
Active Motif Europe
Waterloo Atrium
Drève Richelle 167 – boîte 4BE-1410 Waterloo, Belgium
E-mail: eurotech@activemotif.com
Direct: +32 (0)2 653 0001
Active Motif Japan
Azuma Bldg, 7th Floor
2-21 Ageba-Cho, Shinjuku-Ku
Tokyo, 162-0824, Japan
Active Motif China
787 Kangqiao Road
Building 10, Suite 202
Pudong District
Shanghai, 201315, China

Toll Free: 877 222 9543
Direct: 760 431 1263
Fax: 760 431 1351

Germany Free Phone: 0800 181 99 10
France Free Phone: 0800 90 99 79
UK Free Phone: 0800 169 31 47
Fax: +32 (0)2 653 0050

Direct: +81 3 5225 3638
Fax: +81 3 5261 8733
E-mail: japantech@activemotif.com

Direct: (86)-21-20926090
Hotline: 400-018-8123
E-mail: techchina@activemotif.com

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