Thermo Fisher Scientific PrepFiler™ Forensic DNA Extraction Kit Owner's manual

Brand: Thermo Fisher Scientific

Model: PrepFiler™ Forensic DNA Extraction Kit

Type: Owner's manual

PrepFiler™

Forensic DNA Extraction Kit

User Guide

For Research Use Only. Not for use in diagnostic procedures.

Information in this document is subject to change without notice. Applied Biosystems Inc assumes no responsibility for any errors that may appear in

this document.

APPLIED BIOSYSTEMS DISCLAIMS ALL WARRANTIES WITH RESPECT TO THIS DOCUMENT, EXPRESSED OR IMPLIED, INCLUDING

BUT NOT LIMITED TO THOSE OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE. IN NO EVENT SHALL APPLIED

BIOSYSTEMS BE LIABLE, WHETHER IN CONTRACT, TORT, WARRANTY, OR UNDER ANY STATUTE OR ON ANY OTHER BASIS FOR

SPECIAL, INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING

FROM THIS DOCUMENT, INCLUDING BUT NOT LIMITED TO THE USE THEREOF.

NOTICE TO PURCHASER:

This product or portions thereof is manufactured and sold under license from GE Healthcare under U.S. Patent numbers 5,523,231 and 5,681,946 and

other foreign patents.

End User Terms and Conditions

Acceptance. These terms and conditions shall govern the purchase, use, and transfer and acceptance of the products described in the purchase order,

quotation or invoice, which products are sold and distributed by Applera Corporation to the Buyer/transferee of such products (the “End User”). The

transfer/sale of products to the End User is expressly conditional upon End User’s acceptance of these terms and conditions.

Restrictions on Use. End Users are specifically not authorized to and are forbidden from reselling, transferring or distributing any products either as a

stand alone product or as a component of another product. The right to use the products does not, in and of itself, include or carry any right of the End

User to any GE Healthcare Bio-Sciences Corporation’s technology or intellectual property other than expressly provided herein. End Users may not

attempt to reverse engineer parameters of any of GE Healthcare Bio-Sciences Corporation proprietary products or services.

DISCLAIMER OF WARRANTIES. GE HEALTHCARE BIO-SCIENCES CORPORATION PROVIDES NO WARRANTIES TO END USER

(STATUTORY OR IMPLIED), INCLUDING WITHOUT LIMITATION, AS TO PRODUCT QUALITY, CONDITION, DESCRIPTION,

MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, AND ALL SUCH WARRANTIES ARE HEREBY EXPRESSLY

DISCLAIMED. GE HEALTHCARE BIO-SCIENCES CORPORATION HEREBY EXPRESSLY DISCLAIMS ANY WARRANTY REGARDING

RESULTS OBTAINED THROUGH THE USE OF THE PRODUCTS, INCLUDING WITHOUT LIMITATION ANY CLAIM OF INACCURATE,

INVALID OR INCOMPLETE RESULTS.

Exclusion of Liability. GE Healthcare Bio-Sciences Corporation and its affiliates shall have no liability to an End User, including, without limitation, for

any loss of use or profits, business interruption or any consequential, incidental, special or other indirect damages of any kind, regardless of how caused

and regardless of whether an action in contract, tort, strict product liability or otherwise.

TRADEMARKS:

Applied Biosystems, AB (Design), AmpFlSTR, GeneAmp, GeneMapper, Identifiler, and Quantifiler are registered trademarks and MiniFiler and

PrepFiler are trademarks of Applied Biosystems Inc. or its subsidiaries in the US and/or certain other countries.

All other trademarks are the sole property of their respective owners.

Part Number 4390932 Rev. B

11/2008

Contents

iiiPrepFiler™ Forensic DNA Extraction Kit User Guide

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v

How to use this guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v

Safety information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi

How to obtain more information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . x

How to obtain support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1

PrepFiler™ Forensic DNA Extraction Kit description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Extraction workflow overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

Kit contents and storage conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

Required materials and instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

Chapter 2 Perform Extraction – Standard Protocol . . . . . . . . . . . . . . . . . 9

Standard protocol – sample types and inputs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

Required materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

Tube-handling guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

Magnetic stand guidelines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

Step 1: Prepare reagents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Step 2: Perform lysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

Step 3: Remove substrate from sample lysate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

Step 4: Bind genomic DNA to magnetic particles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

Step 5: Wash bound DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

Step 6: Elute DNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

Chapter 3 Perform Extraction – Supplementary Protocols . . . . . . . . . . 23

Supplementary protocols for additional sample types and inputs . . . . . . . . . . . . . . . . . 24

Blood/soil mixture protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

Epithelial- and sperm-cell fraction protocols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

Hair protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Nail cuttings protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

Paraffin-embedded tissue protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

Tooth protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

Large-sample protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

Contents

PrepFiler™ Forensic DNA Extraction Kit User Guide

Chapter 4 Experiments and Results . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

Sensitivity study (SWGDAM Guideline 2.3) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

Stability study (SWGDAM Guideline 2.4) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

Reproducibility study (SWGDAM Guideline 2.5) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48

Case-type sample study (SWGDAM Guideline 2.6) . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50

Contamination study (SWGDAM Guideline 3.6) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52

Correlation study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53

Correlation with STR analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 57

Appendix A Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

Appendix B Repurification Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

vPrepFiler™ Forensic DNA Extraction Kit User Guide

Preface

How to use this guide

Purpose of this

guide

This guide provides step-by-step instructions for using the PrepFiler™ Forensic DNA

Extraction Kit to extract and isolate genomic DNA from forensic samples, and a

description of the experiments performed by Applied Biosystems to evaluate the

PrepFiler Forensic DNA Extraction Kit.

Audience This guide is intended for scientists who isolate DNA from forensic samples for the

purpose of quantitation and Short Tandem Repeat (STR) analysis.

Tex t conv entio ns This guide uses the following conventions:

•Bold text indicates user action. For example:

Add 15 µL of PrepFiler™ Magnetic Particles to the tube.

•Italic text indicates new or important words and is also used for emphasis.

For example:

Before analyzing, always prepare fresh matrix.

User attention

words

Two user attention words appear in Applied Biosystems user documentation. Each

word implies a particular level of observation or action as described below:

Note: – Provides information that may be of interest or help but is not critical to the

use of the product.

IMPORTANT! – Provides information that is necessary for proper instrument

operation, accurate chemistry kit use, or safe use of a chemical.

Examples of the user attention words appear below:

Note: You can also use a heat block to incubate the samples.

IMPORTANT! Do not disturb the magnetic particles pellet.

Preface

PrepFiler™ Forensic DNA Extraction Kit User Guide

Safety information

Safety alert

words

Four safety alert words appear in Applied Biosystems user documentation at points

in the document where you need to be aware of relevant hazards. Each alert

word—IMPORTANT, CAUTION, WARNING, DANGER—implies a particular

level of observation or action, as defined below.

Definitions

IMPORTANT! – Indicates information that is necessary for proper instrument

operation, accurate chemistry kit use, or safe use of a chemical.

CAUTION! – Indicates a potentially hazardous situation that, if not avoided,

may result in minor or moderate injury. It may also be used to alert against

unsafe practices.

WARNING! – Indicates a potentially hazardous situation that, if not avoided,

could result in death or serious injury.

DANGER! – Indicates an imminently hazardous situation that, if not avoided,

will result in death or serious injury. This signal word is to be limited to the

most extreme situations.

Chemical hazard

warning

WARNING! CHEMICAL HAZARD. Some of the chemicals used with

Applied Biosystems instruments and protocols are potentially hazardous and

can cause injury, illness, or death.

Chemical safety

guidelines

To minimize the hazards of chemicals:

• Read and understand the Material Safety Data Sheets (MSDSs) provided by the

chemical manufacturer before you store, handle, or work with any chemicals or

hazardous materials. (See “About MSDS” on page vii.)

• Minimize contact with chemicals. Wear appropriate personal protective

equipment when handling chemicals (for example, safety glasses, gloves, or

protective clothing). For additional safety guidelines, consult the MSDS.

• Minimize the inhalation of chemicals. Do not leave chemical containers open.

Use only with adequate ventilation (for example, fume hood). For additional

safety guidelines, consult the MSDS.

• Check regularly for chemical leaks or spills. If a leak or spill occurs, follow the

manufacturer’s cleanup procedures as recommended in the MSDS.

• Comply with all local, state/provincial, or national laws and regulations related

to chemical storage, handling, and disposal.

vii

Safety information

PrepFiler™ Forensic DNA Extraction Kit User Guide

About MSDS Chemical manufacturers supply current Material Safety Data Sheets (MSDSs) with

shipments of hazardous chemicals to new customers. They also provide MSDSs with

the first shipment of a hazardous chemical to a customer after an MSDS has been

updated. MSDSs provide the safety information you need to store, handle, transport,

and dispose of the chemicals safely.

Each time you receive a new MSDS packaged with a hazardous chemical, be sure to

replace the appropriate MSDS in your files.

Obtaining

MSDSs

The MSDSs for any chemicals supplied by Applied Biosystems or Ambion are

available to you free 24 hours a day.

IMPORTANT! For the MSDSs of chemicals not distributed by Applied Biosystems

or Ambion contact the chemical manufacturer.

To obtain MSDSs:

1. Go to www.appliedbiosystems.com, click Support, then click MSDS.

2. In the Keyword Search field, enter the chemical name, product name, MSDS

part number, or other information that appears in the MSDS of interest. Select

the language of your choice, then click Search.

3. Find the document of interest, right-click the document title, then select any of

the following:

•Open – To view the document

•Print Target – To print the document

•Save Target As – To download a PDF version of the document to a

destination that you choose

IMPORTANT! For the MSDSs of chemicals not distributed by Applied Biosystems

or Ambion contact the chemical manufacturer.

Chemical waste

hazards

CAUTION! HAZARDOUS WASTE. Refer to Material Safety Data Sheets

and local regulations for handling and disposal.

WARNING! CHEMICAL WASTE HAZARD. Wastes produced by Applied

Biosystems instruments are potentially hazardous and can cause injury, illness,

or death.

WARNING! CHEMICAL STORAGE HAZARD. Never collect or store

waste in a glass container because of the risk of breaking or shattering.

Reagent and waste bottles can crack and leak. Each waste bottle should be

secured in a low-density polyethylene safety container with the cover fastened

and the handles locked in the upright position. Wear appropriate eyewear,

clothing, and gloves when handling reagent and waste bottles.

viii

Preface

PrepFiler™ Forensic DNA Extraction Kit User Guide

Chemical waste

safety guidelines

To minimize the hazards of chemical waste:

• Read and understand the Material Safety Data Sheets (MSDSs) provided by the

manufacturers of the chemicals in the waste container before you store, handle,

or dispose of chemical waste.

• Provide primary and secondary waste containers. (A primary waste container

holds the immediate waste. A secondary container contains spills or leaks from

the primary container. Both containers must be compatible with the waste

material and meet federal, state, and local requirements for container storage.)

• Minimize contact with chemicals. Wear appropriate personal protective

equipment when handling chemicals (for example, safety glasses, gloves, or

protective clothing). For additional safety guidelines, consult the MSDS.

• Minimize the inhalation of chemicals. Do not leave chemical containers open.

Use only with adequate ventilation (for example, fume hood). For additional

safety guidelines, consult the MSDS.

• Handle chemical wastes in a fume hood.

• After emptying the waste container, seal it with the cap provided.

• Dispose of the contents of the waste tray and waste bottle in accordance with

good laboratory practices and local, state/provincial, or national environmental

and health regulations.

Waste disposal If potentially hazardous waste is generated when you operate the instrument, you

must:

• Characterize (by analysis if necessary) the waste generated by the particular

applications, reagents, and substrates used in your laboratory.

• Ensure the health and safety of all personnel in your laboratory.

• Ensure that the instrument waste is stored, transferred, transported, and disposed

of according to all local, state/provincial, and/or national regulations.

IMPORTANT! Radioactive or biohazardous materials may require special handling,

and disposal limitations may apply.

Safety information

PrepFiler™ Forensic DNA Extraction Kit User Guide

Biological hazard

safety WARNING! BIOHAZARD. Biological samples such as tissues, body fluids,

infectious agents, and blood of humans and other animals have the potential to

transmit infectious diseases. Follow all applicable local, state/provincial,

and/or national regulations. Wear appropriate protective equipment, which

includes but is not limited to: protective eyewear, face shield, clothing/lab coat,

and gloves. All work should be conducted in properly equipped facilities using

the appropriate safety equipment (for example, physical containment devices).

Individuals should be trained according to applicable regulatory and

company/institution requirements before working with potentially infectious

materials. Read and follow the applicable guidelines and/or regulatory

requirements in the following:

• U.S. Department of Health and Human Services guidelines published in

Biosafety in Microbiological and Biomedical Laboratories (stock no. 017-040-

00547-4; bmbl.od.nih.gov)

• Occupational Safety and Health Standards, Bloodborne Pathogens (29

CFR§1910.1030; www.access.gpo.gov/

nara/cfr/waisidx_01/29cfr1910a_01.html).

• Your company’s/institution’s Biosafety Program protocols for working

with/handling potentially infectious materials.

Additional information about biohazard guidelines is available at:

www.cdc.gov

Preface

PrepFiler™ Forensic DNA Extraction Kit User Guide

How to obtain more information

documentation

The following related document is available from Applied Biosystems:

•PrepFiler™ Forensic DNA Extraction Kit Quick Reference Card

(PN 4393918) – Provides brief, step-by-step procedures for isolating genomic

DNA. It is designed to be used as a reference in the laboratory after you become

familiar with the content in the User Guide.

For additional documentation, see “How to obtain support” on page xi.

Send us your

comments

Applied Biosystems welcomes your comments and suggestions for improving its

user documents. You can e-mail your comments to:

techpubs@appliedbiosystems.com

The e-mail address above is only for submitting comments and suggestions relating

to documentation. To order documents, download PDF files, or for help with a

technical question, go to http://www.appliedbiosystems.com, then click the link for

Support. (See “How to obtain support” on page xi).

How to obtain support

PrepFiler™ Forensic DNA Extraction Kit User Guide

How to obtain support

For HID support:

• In North America – send an email to

HIDTechSupport@appliedbiosystems.com, or

call 888-821-4443 option 1.

• Outside North America – contact your local support office.

For the latest services and support information for all locations, go to

http://www.appliedbiosystems.com, then click the link for Support. At the Support

page, you can:

• Access worldwide telephone and fax numbers to contact Applied Biosystems

Technical Support and Sales facilities

• Search through frequently asked questions (FAQs)

• Submit a question directly to Technical Support

• Order Applied Biosystems user documents, MSDSs, certificates of analysis,

and other related documents

• Download PDF documents

• Obtain information about customer training

• Download software updates and patches

xii

Preface

PrepFiler™ Forensic DNA Extraction Kit User Guide

1PrepFiler™ Forensic DNA Extraction Kit User Guide

Introduction

This chapter covers:

PrepFiler™ Forensic DNA Extraction Kit description . . . . . . . . . . . . . . . . . . . . . . . 2

Extraction workflow overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

Kit contents and storage conditions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

Required materials and instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

Chapter 1 Introduction

PrepFiler™ Forensic DNA Extraction Kit User Guide

PrepFiler™ Forensic DNA Extraction Kit description

The PrepFiler™ Forensic DNA Extraction Kit contains Applied Biosystems reagents

optimized for use in:

• Performing cell lysis

• Binding genomic DNA to magnetic particles

• Removing PCR inhibitors

• Eluting concentrated purified DNA

The kit can be used with one standard protocol to extract and isolate DNA from most

forensic sample types, including body fluids, stains and swabs of body fluids, and

small tissue samples. The approximate yield from 1 µL of blood that contains 4,000

to 11,000 nucleated blood cells is 25 to 65 ng of DNA. The kit is appropriate for use

with samples containing potential inhibitors of the polymerase chain reaction (PCR).

The extracted DNA is compatible for use in quantitation using the Quantifiler®

Human, Quantifiler® Y Human Male, and Quantifiler® Duo DNA Quantification

Kits, and for use in STR amplification using the AmpFlSTR® PCR Amplification

kits.

Extraction workflow overview

PrepFiler™ Forensic DNA Extraction Kit User Guide

Extraction workflow overview

This user guide contains a standard extraction protocol that is appropriate for most

forensic sample types (see Chapter 2 on page 9), and supplementary protocols that

cover additional sample types such as hair, nails, and teeth (see Chapter 3 on

page 23). The standard protocol workflow is shown below. The supplementary

protocols follow similar workflows, with some differences due to variations in

sample preparation and lysis.

Perform Cell Lysis

Remove Substrate

Bind DNA

Elute DNA

Wash DNA

Prepare Reagents

Standard Protocol

(Chapter 2)

Chapter 1 Introduction

PrepFiler™ Forensic DNA Extraction Kit User Guide

Kit contents and storage conditions

Kit contents Each PrepFiler™ Forensic DNA Extraction Kit contains materials sufficient to

perform 100 extractions using the standard protocol (300 µL PrepFiler™ Lysis Buffer

per extraction). The contents of the kits are described in Table 1.

Table 1 Materials provided with the PrepFiler™ Forensic DNA Extraction Kits

(PNs 4392852 and 4392353)

Reagent Description

Included with

Part Number

4392852‡

‡ The kit includes filter columns and spin tubes for sample substrate removal. Extra spin tubes are included for use in the lysis and elution

steps.

Included with

Part Number

4392353

PrepFiler™ Lysis Buffer One bottle, 50 mL ✔✔

PrepFiler™ Isopropanol One empty 60 mL bottle (user provides

isopropanol)

✔✔

PrepFiler™ Magnetic Particles One tube, 1.5 mL ✔✔

PrepFiler™ Wash Buffer

Concentrate

Two 125-mL bottles, each containing

26 mL of concentrate (user adds 74 mL

of ethanol to each bottle)

✔✔

PrepFiler™ Elution Buffer One bottle, 12.5 mL ✔✔

PrepFiler™ Filter Columns 100 filter columns ✔no

PrepFiler™ Spin Tubes 300 spin tubes ✔no

Kit contents and storage conditions

PrepFiler™ Forensic DNA Extraction Kit User Guide

Storage

conditions

The kits are shipped at room temperature.

IMPORTANT! When you receive a kit, immediately store the PrepFiler™ Magnetic

Particles tube at 4 to 8 °C. Do not freeze the PrepFiler Magnetic Particles tube.

Note: The PrepFiler Magnetic Particles may be shipped in a separate container.

Table 2 PrepFiler™ Forensic DNA Extraction Kit storage conditions

Reagent Storage Conditions

Magnetic Particles • 4 to 8 °C upon receiving the kit

• After first use, store at room temperature

(18 to 25 °C) for up to 3 months or at 4 °C

for longer periods up to the expiration date

Lysis Buffer

Isopropanol

Wash Buffer Concentrate

Prepared Wash Buffer

Elution Buffer

Room temperature (18 to 25 °C)

Chapter 1 Introduction

PrepFiler™ Forensic DNA Extraction Kit User Guide

Required materials and instruments

About thermal

shakers

A thermal shaker was used in the validation studies described in Chapter 4 and is

recommended for DNA extraction using the PrepFiler™ Forensic DNA Extraction

Kit.

You can use a heat block (following the heat block instructions provided in the

protocols) instead of a thermal shaker, however the kit performance has not been

extensively tested under these conditions, and DNA yield may be lower than would

be obtained using a thermal shaker.

Each lab should perform studies to validate the performance of the PrepFiler kit

when using methodologies other than a thermal shaker.

Required

materials and

instruments

Table 3 lists materials and instruments that are required in addition to the reagents

and materials supplied with the PrepFiler™ Forensic DNA Extraction Kit.

Table 3 Required materials and instruments

Protocol Step Material or Instrument Source‡

All Common laboratory equipment such as pipettors,

aerosol-resistant micropipette tips, and a microcentrifuge

Major laboratory supplier

All Vortexer (a variable-speed vortexer is recommended) Major laboratory supplier

Lysis DL-Dithiothreitol [Molecular biology grade; ≥98% (TLC),

≥99% (titration)]

Sigma-Aldrich

www.sigmaaldrich.com

(Part Number D9779)

Lysis RNase-free Microfuge Tubes (1.5 mL), certified DNase- and

RNase-free

Note: 1.5-mL tubes are required for use with the

reagent-only version of the PrepFiler™ kit (PN 4392353),

which does not include PrepFiler™ Filter Columns and Spin

Tubes.

Applied Biosystems

(PN AM12400) or equivalent

Note: If you use an equivalent

tube, select tubes that allow you

to observe the tube contents.

Lysis and elution (Recommended) Eppendorf Thermomixer R or similar

thermal shaker

Heat block (see “About thermal shakers” above; a heat

block also requires more hands-on processing time than a

thermal shaker)

Eppendorf North America

www.eppendorfna.com

Major laboratory supplier

Substrate removal Laboratory centrifuge capable of 16,110 x g Major laboratory supplier

Binding Isopropanol (2-Propanol, ACS reagent grade, ≥99.5%)

Note: Open a new bottle when preparing the PrepFiler™

Isopropanol bottle.

Sigma-Aldrich

www.sigmaaldrich.com

(Part Number 190764)

Required materials and instruments

PrepFiler™ Forensic DNA Extraction Kit User Guide

Binding Laboratory shaker for microcentrifuge tubes

Vortexer with adaptor for microcentrifuge tubes such as a

VWR Microtube Holder (holds forty-eight 0.25-2.0 mL

microcentrifuge tubes)

Major laboratory supplier

VRW

www.vwr.com

(Catalog number 12620-876)

Wash Ethanol (Molecular biology grade; 95% or 190 proof)

Note: Open a new bottle when preparing the PrepFiler™

Wash Buffer solution.

Sigma-Aldrich

www.sigmaaldrich.com

(Part Number E7148)

Wash and elution 6-Tube Magnetic Stand Applied Biosystems

(PN AM10055)

‡ Recommended sources. Equivalent materials from other suppliers can be used after appropriate validation studies by the user

laboratory.

Table 3 Required materials and instruments (continued)

Protocol Step Material or Instrument Source‡

Chapter 1 Introduction

PrepFiler™ Forensic DNA Extraction Kit User Guide

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Thermo Fisher Scientific PrepFiler™ Forensic DNA Extraction Kit Owner's manual

Thermo Fisher Scientific PrepFiler™ Forensic DNA Extraction Kit Owner's manual

Thermo Fisher Scientific PrepFiler™ Forensic DNA Extraction Kit Owner's manual

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