instructions

GFX PCR DNAand Gel BandPurification Kit

product code:

27-9602-01

WarningFor research use only.Not recommended orintended for the diagnosisof disease in humans or animals.Do not use internally orexternally in humans or animals.

Harmful

i 74003977 Ed. AA

● p2

HandlingStorageStore at ambienttemperature.

Page finderComponents of the kit . . . . . . . . . . . . . . . . . . . . . 3

Quality control . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

Materials not supplied . . . . . . . . . . . . . . . . . . . . . 3

World Wide Web address. . . . . . . . . . . . . . . . . . . 4

Safety warnings and precautions . . . . . . . . . . . . . 4

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

ProtocolsEssential preliminaries . . . . . . . . . . . . . . . . . . . . . 71. Purification of DNA from solution . . . . . . . . . . 82. Purification of DNA from gel bands . . . . . . . . . 9

Appendixes1. Protocol for MicroPlex 24 Vacuum. . . . . . . . . 112. Protocol for MicroPlex 24 (Spin) . . . . . . . . . . 15

Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . 19

References . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

Companion products . . . . . . . . . . . . . . . . . . . . . 22

Licenses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

Trademarks . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

Components of the kit

The following components are included inthis product:

Capture buffer Buffered solution containingacetate and chaotrope.

GFX columns MicroSpin™ columnspre-packed with a glassfiber matrix.

Collection tubes 2 ml capless microcentrifugetubes.

Wash buffer* Tris-EDTA buffer (10 mMTris-HCl pH 8.0, 1mM EDTA).Add absolute ethanol to afinal concentration of 80%before use as indicated inthe note below.

*The wash buffer requires dilution before use. Tothe bottle containing wash buffer, add 48 ml ofabsolute ethanol. Mix by inversion. After dilutingthis component, indicate on the label that thisstep has been completed. When not in use, storethe wash buffer-ethanol mixture AIRTIGHT atambient temperatures.

Quality controlGFX™ PCR DNA and GelBand Purification Kit istested for the ability toquantitatively isolate a910 base-pair PCR productfrom solution and from anagarose gel band.

Materials notsuppliedReagents

• Absolute Ethanol

• Elution Buffer—10 mMTris-HCl, pH 8.0 or TEbuffer (10 mM Tris-HCl,pH 8.0, 1 mM EDTA) orautoclaved double-dis-tilled water.

Equipment

• Microcentrifuge—thataccomodates 1.5 mlmicrocentrifuge tubes.

• Tubes—1.5 mlmicrocentrifuge tubes

• Scalpel or razor blade

• Incubator or waterbath—60 ˚C

● p3

● p4

World Wide Web address

http://www.gehealthcare.com/lifesciences

Visit the GE Healthcare home page for regularly updated prod-uct information.

Safety warnings and precautions

Warning: For research use only. Not recommended or intended fordiagnosis of disease in humans or animals. Do not use internally orexternally in humans or animals.

All chemicals should be considered as potentially hazardous. Onlypersons trained in laboratory techniques and familiar with the principlesof good laboratory practice should handle these products. Suitableprotective clothing such as laboratory overalls, safety glasses, andgloves should be worn. Care should be taken to avoid contact withskin or eyes; if contact should occur, wash immediately with water(see Material Safety Data Sheet for specific recommendations).

Warning: This protocol requires the use of ethanol and formamide.Gel reagents may contain acrylamide, a neurotoxin and suspectedcarcinogen. Please follow the manufacturer’s Material Safety DataSheet regarding safe handling and use of these materials.

● p5

Introduction

GFX PCR DNA and Gel Band Purification Kit uses a chaotropic agentthat denatures protein, dissolves agarose, and promotes the binding ofdouble-stranded DNA (100 base-pairs to 48 kilobase-pairs) to a glassfiber matrix (2, 3). Once the DNA is “captured”, proteins and saltcontaminants are washed away, and the purified DNA is eluted in alow ionic strength buffer (TE, Tris-HCl or water). DNA samples arerecovered in a concentrated form by eluting with as little as 10 µl ofbuffer or water.

GFX PCR DNA and Gel Band Purification Kit can be used to purifyDNA (e.g., PCR products, restriction fragments) from solution andfrom both TAE and TBE agarose gel bands. Typical recoveries are>80% from solution and > 60% from gel bands. GFX purificationof PCR products from solution removes 99.5% of primers and freenucleotides and can be performed in less than 5 minutes. DNA purifica-tion from a gel band can be completed in just 15 minutes. PurifiedDNA can be used directly without an alcohol precipitation in a varietyof applications including PCR amplification, restriction digest analysisand subcloning.

To efficiently process 24, 48 or 96 samples simultaneously, the kit canbe used with MicroPlex™ 24 Vacuum or MicroPlex 24 (see Appendixes1 and 2, respectively). Use with MicroPlex 24 Vacuum can result intime savings of >20% for 48 samples and >40% for 96 samples ascompared with microfuge based purification.

● p6

The basic protocol for the GFX PCR DNA and Gel Band PurificationKit involves the following steps:

Step Comments Component

Denaturation/ Proteins are denatured. For gel band Capture buffersolubilization purification, agarose is dissolved.

Capture The sample is passed through the GFX columnGFX column to capture the DNAonto the glass fiber matrix.

Washing/ Matrix-bound DNA is washed with Wash bufferDrying an ethanolic buffer to remove salts

and other contaminants. This stepalso dries the matrix prior to elution.

Elution The purified DNA is eluted from the Not suppliedGFX column in low ionic strengthbuffer (10 mM Tris-HCl pH 8.0,TE pH 8.0 or water).

ProtocolsEssential Preliminaries

To elute the purified plasmid DNA, we recommend using either 10 mMTris-HCl pH 8.0, TE buffer (10 mM Tris-HCl pH 8.0, 1 mM EDTA) or auto-claved double-distilled water.

Prior to the first use, the wash buffer must be diluted (see page 3).

After diluting, indicate on the label that this step has been completed.

Purification from solution or from gel bandsProcedures 1 and 2 provide protocols for purifying PCR products, restric-tion fragments and plasmid DNA from solution or from agarose gelbands, respectively. Double-stranded DNA can be purified from solutionvolumes of up to 100 µl and from gel slices of up to 300 mg. Note thatno modifications are required for purification from gels run in borate-based buffers (e.g., TBE).

Eluting the DNA in a concentrated formTo recover DNA in a concentrated form, use elution volumes that areless than the volume of the sample being purified. For optimal recoveryuse volumes ≥ 50 µl. Using elution volumes less than 50 µl will reducethe yield. For example DNA eluted in 30 µl will concentrate the recov-ered DNA approximately 1.5 Fold with a loss of 20–30% as compared toa 50 µl elution volume. An elution with 10 µl will concentrate the DNArecovered approximately 2.5 Fold with a loss of 60–70%.

● p7

➊ Purification of DNA from solution

The maximum volume of PCR reaction/DNA solution that can beprocessed with the following protocol is 100 µl.

1.1 Place one GFX column in a collection tube for each purificationto be performed.

1.2 Add 500 µl of capture buffer to the GFX column.

1.3 Transfer the DNA solution (up to 100 µl) to the GFX column.If purifying a PCR sample, avoid transferring mineral oil tothe column.

1.4 Mix thoroughly by pipetting the sample up and down 4–6 times.

1.5 Centrifuge in a microcentrifuge at full speed for 30 s.

1.6 Discard the flow-through by emptying the collection tube. Placethe GFX column back inside the collection tube.

1.7 Add 500 µl of wash buffer to the column. Centrifuge at full speedfor 30 s.

1.8 Discard the collection tube and transfer the GFX column to afresh 1.5 ml microcentrifuge tube (i.e. NOT a collection tube).

1.9 Apply 50 µl of elution buffer (10 mM Tris-HCl pH 8.0, TE pH 8.0or autoclaved double-distilled water) directly to the top of theglass fiber matrix in the GFX Column.

To elute DNA in a more concentrated form reduce the elutionvolume to no less than 10 µl.

Note: Using low elution volumes to concentrate the DNA samplewill reduce recovery (see page 7).

● 8

1.10 Incubate the sample at room temperature for 1 min.

1.11 Centrifuge at full speed for 1 min to recover the purified DNA.

Note: For 50 µl elutions, the actual volume of sample recoveredwill range from 40 to 50 µl. For a 10 µl elution, the recoveredvolume will range from 5 to 7 µl.

➋ Purification of DNA from gel bands

The maximum weight of gel slice that can be processed with thefollowing protocol is 300 mg; the column can accommodate amaximum volume of 600 µl (i.e., 300 mg of gel slice and 300 µl ofcapture buffer).

2.1 Weigh an empty 1.5 ml microcentrifuge tube to the nearest10 mg and record the weight.

2.2 Using a clean razor blade or scalpel, excise the slice of agarosecontaining the DNA band to be purified. Cut as close to the DNAband as possible. Cut the slice into several smaller pieces andtransfer them to the pre-weighed 1.5 ml microcentrifuge tube.

2.3 Weigh the tube containing the agarose slice to the nearest10 mg, and subtract the weight of the empty tube to determinethe weight of the slice.

2.4 To the gel slice add 10 µl of capture buffer for each 10 mg of gelslice (maximum column capacity is 300 µl of capture bufferadded to a 300 mg gel slice).

2.5 Close the tube and mix by vortexing vigorously. Incubate at 60 ˚Cuntil the agarose is completely dissolved (5–15 min).

● p9

2.6 During the incubation, place one GFX column in a collection tubefor each purification to be performed.

2.7 After the agarose is completely dissolved, centrifuge briefly tocollect the sample at the bottom of the tube.

2.8 Transfer the sample to the GFX column. Incubate at room tem-perature for 1 min.

2.9 Centrifuge in a microcentrifuge at full speed for 30 s.

2.10 Discard the flow-through by emptying the collection tube. Placethe GFX column back inside the collection tube.

2.11 Add 500 µl of wash buffer to the column. Centrifuge at full speedfor 30 s.

2.12 Discard the collection t and transfer the GFX column to a fresh1.5 ml microcentrifuge tube (i.e. NOT a collection tube).

2.13 Apply 50 µl of elution buffer (10 mM Tris-HCl pH 8.0, TE pH 8.0or autoclaved double-distilled water) directly to the top of theglass fiber matrix in the GFX column.

To elute DNA in a more concentrated form reduce the elutionvolume to no less than 10 µl.

Note: Using low elution volumes to concentrate the DNA samplewill reduce recovery (see page 7).

2.14 Incubate the sample at room temperature for 1 min.

2.15 Centrifuge at full speed for 1 min to recover the purified DNA.

Note: For 50 µl elutions, the actual volume of sample recoveredwill range from 40 to 50 µl. For a 10 µl elution, the recoveredvolume will range from 5 to 7 µl.

● p10

Appendix 1:Protocol for MicroPlex 24 Vacuum

Either MicroPlex 24 Vacuum or MicroPlex 24 may be used to effi-ciently process large numbers of samples simultaneously. If a vacuumsource capable of generating ~200 mm Hg (e.g. a house vacuum) isavailable, we recommend that MicroPlex 24 Vacuum be used for highthroughput purification. If a tabletop centrifuge and the appropriatemicrotiter plate rotor is available, MicroPlex 24 may be used.

Using the MicroPlex 24 systems will result in significant time savings.For example, using MicroPlex 24 Vacuum can result in time savings of>20% for 48 samples and >40% for 96 samples as compared withmicrofuge-based purification. For either MicroPlex procedure, DNArecovery will be approximately 30% to 40% lower than that achievedwith standard kit protocols.

To use the MicroPlex 24 Vacuum, follow the “MicroPlex 24 VacuumAssembly” steps below and then go to either “Option A: Purification ofDNA from solution” (page 12) or “Option B: Purification of DNAfrom gel bands” (page 12) depending on the sample type.

MicroPlex 24 Vacuum assembly

1. Use the “Vacuum Applications” instructions provided with theMicroPlex 24 to assemble the manifold and connect it to thevacuum source.

2. Place the required number of GFX columns in MicroPlex 24manifold.

● p11

3. Fill any unused holes in the manifold with plugs included withMicroPlex 24 Vacuum.

Note: Vacuum should NOT be applied to the manifold until ALL ofthe samples have been transferred to the columns and incubated.

4. Make sure that the vacuum line stopcock is in the closed position(i.e. perpendicular to the vacuum tubing) and that the manifold isplaced squarely on the gasket and collection tray. This is necessary toensure proper application of the vacuum.

5. Proceed to “Option A” if purifying DNA from solution, or alterna-tively use “Option B” if purifying from gel bands (see below).

Option A: Purification of DNA from solution

1. Add 500 µl of capture buffer to each GFX column.

2. Transfer the DNA solution (up to 100 µl) to the GFX columns in theMicroPlex 24. If purifying a PCR sample, avoid transferring mineraloil to the column.

3. Proceed to “DNA binding and recovery” page 13.

Option B: Purification of DNA from gel bands

1. Weigh an empty 1.5 ml microcentrifuge tube to the nearest 10 mgand record the weight.

2. Using a clean razor blade or scalpel, excise the slice of agarose con-taining the DNA band to be purified. Cut as close to the DNA bandas possible. Cut the slice into several smaller pieces and transfer themto the pre-weighed 1.5 ml microcentrifuge tube.

3. Weigh the tube containing the agarose slice to the nearest 10 mg,and subtract the weight of the empty tube to determine the weight ofthe slice.

● p12

4. To the gel slice add 10 µl of capture buffer for each 10 mg of gel slice(maximum column capacity is 300 µl of capture buffer added to a300 mg gel slice).

5. Close the tube and mix by vortexing vigorously. Incubate at 60 ˚Cuntil the agarose is completely dissolved (5–15 minutes).

6. After the agarose is completely dissolved, centrifuge briefly to collectthe sample at the bottom of the tube.

7. Transfer the sample (up to 600 µl) to the GFX columns in theMicroPlex 24. Incubate at room temperature for 1 min.

8. Proceed to “DNA binding and recovery,” immediately below.

DNA binding and recovery

1. Turn the vacuum supply (e.g. house vacuum) on at the source. Turnthe stopcock in the vacuum tubing assembly to the open position(i.e. parallel to the tubing). After the samples have been drawnthrough all of the columns and into the collection tray, turn thestopcock to the closed position, leaving the vacuum supply on atthe source.

2. Add 500 µl of wash buffer to each column. Turn the stopcock to theopen position. Allow the solution to be drawn through each columnand continue to apply the vacuum for 10 min to remove residualbuffer and dry the matrix.

Note: If using 2, 3 or 4 manifolds connected to the same vacuumsource in parallel, dry 15–20 min. Avoid over-drying (>30 min); DNAmay become irreversibly bound to the glass fiber matrix.

● p13

3. Turn the stopcock to the closed position. Allow the vacuum pressurehas fully dissipated, remove the manifold from the collection tray andplace it on a clean paper towel. (The manifold should separate easilyfrom the collection tray.) Discard the used collection tray and savethe gasket.

4. For short-term storage, purified plasmid DNA can be collected usingMicroPlex 24 Vacuum. (If desired, the trays containing the final sam-ples can be covered with sealing tape from Costar, catalog no. 3095).Transfer the gasket and manifold to a new collection tray. Reassemblethe MicroPlex 24 Vacuum system with the new collection tray.Proceed to the next step.

Alternatively, the columns can be transferred to 1.5 ml microcen-trifuge tubes for collection of samples using a microcentrifuge. Thiseliminates the need to transfer samples from the collection tray totubes for long term storage.

5. Apply 125 µl of elution buffer (10 mM Tris-HCl pH 8.0 or TE pH 8.0or autoclaved double-distilled water) directly to the top of the glassfiber matrix in the GFX column.

6. Incubate the sample at room temperature for 1 min.

7. To collect the purified DNA using MicroPlex 24 Vacuum, turn thestopcock to the open position. After all the samples have been drawnthrough the columns and into the collection tray, turn the stopcockto the closed position. Allow the vacuum pressure to dissipate for10–15 seconds and separate the collection tray (which contains thepurified plasmid DNA) from the manifold as previously described.If desired, the tray containing the final samples can be covered withsealing tape from Costar (catalog no. 3095).

● p14

8. Alternatively, if using 1.5 ml microcentrifuge tubes, recover the DNAby centrifugation at full speed for 1 min in a microcentrifuge.

Note: The recovered elution volume will be 40–60% of the appliedvolume for the MicroPlex 24 Vacuum method of sample recovery.DNA recovery will be approximately 30% to 40% lower than thatfrom the microfuge procedure.

Appendix 2:Protocol for MicroPlex 24 (Spin)

Either MicroPlex 24 Vacuum or MicroPlex 24 may be used to effi-ciently process large numbers of samples simultaneously. If a vacuumsource capable of generating ~200 mm Hg (e.g. a house vacuum) isavailable, we recommend that MicroPlex 24 Vacuum be used for highthroughput purification. If a tabletop centrifuge and the appropriatemicrotiter plate rotor is available, MicroPlex 24 may be used.

Using the MicroPlex 24 systems will result in significant time savings.For example, using the MicroPlex 24 Vacuum can result in time savingsof >20% for 48 samples and >40% for 96 samples as compared tomicrofuge based methods. For either MicroPlex procedure, DNA recov-ery will be approximately 30% to 40% lower than with the microfugeprocedures.

To use the MicroPlex 24, follow the “MicroPlex 24 assembly” stepsand then go to either “Option A: Purification of DNA from solution”(page 16) or “Option B: Purification of DNA from gel bands”(page 16) depending on the sample type.

● p15

MicroPlex 24 Assembly

1. Use the “Spin Applications” instructions provided with theMicroPlex 24 to assemble the manifold and V-bottom collection tray.

2. Place the required number of GFX columns in the MicroPlex 24manifold.

3. Proceed to page 16 and use “Option A” if purifying DNA from solu-tion, or alternatively use “Option B” if purifying from gel bands.

Option A: Purification of DNA from solution

1. Add 500 µl of capture buffer to each GFX column.

2. Transfer the DNA solution (up to 100 µl) to the GFX columns in theMicroPlex 24 manifold. If purifying a PCR sample, avoid transferringmineral oil to the column.

3. Proceed to “DNA binding and recovery” page 17.

Option B: Purification of DNA from gel bands

1. Weigh an empty 1.5 ml microcentrifuge tube to the nearest 10 mgand record the weight.

2. Using a clean razor blade or scalpel, excise the slice of agarose con-taining the DNA band to be purified. Cut as close to the DNA bandas possible. Cut the slice into several smaller pieces and transfer themto the pre-weighed 1.5 ml microcentrifuge tube.

3. Weigh the tube containing the agarose slice to the nearest 10 mg,and subtract the weight of the empty tube to determine the weight ofthe slice.

● p16

4. To the gel slice add 10 µl of capture buffer for each 10 mg of gel slice(maximum column capacity is 300 µl of capture buffer added to a300 mg gel slice).

5. Close the tube and mix by vortexing vigorously. Incubate at 60˚ Cuntil the agarose is completely dissolved (5-15 min).

6. After the agarose is completely dissolved, centrifuge briefly to collectthe sample at the bottom of the tube.

7. Transfer the sample (up to 600 µl) to the GFX columns in theMicroPlex 24. Incubate at room temperature for 1 min.

8. Proceed to “DNA binding and recovery” page 17.

DNA binding and recovery

1. Centrifuge the MicroPlex 24 unit for 2 min at 1 000 × g (a minimumforce of 700 × g is needed).

Note: The appropriate centrifugation speed for a specific rotor can becalculated from the following formula.

RCF = (1.12)(r)(rpm/1 000)2

where RCF = relative centrifugal force;r = radius in mm measured from the center of the spindle to the bot-tom of the rotor bucket; and rpm = revolutions per minute.

2. Add 500 µl of wash buffer to each column. Centrifuge theMicroPlex 24 unit for 10 min.

● p17

3. For short-term storage, purified DNA can be collected usingMicroPlex 24 and a collection tray. If desired, the trays containingthe final samples can be covered with sealing tape from Costar (cata-log no. 3095). To collect samples using MicroPlex 24, remove themanifold from the collection tray and place it on a clean paper towel.Reassemble the MicroPlex 24 unit with a fresh collection tray andproceed to the next step. Discard the used collection tray.

Alternatively, the columns can be transferred to 1.5 ml micro-centrifuge tubes for collection of samples using a microcentrifuge.This eliminates the need to transfer samples from the collection trayto tubes for long-term storage.

4. Apply 100 µl of elution buffer (10 mM Tris-HCl pH 8.0 or TE pH8.0 or autoclaved double-distilled water) directly to the top of theglass fiber matrix in the GFX column.

5. Incubate the samples at room temperature for 1 min.

6. To collect the purified DNA, centrifuge the MicroPlex 24 for 2 minaccording to the instructions supplied with the unit. Alternatively, ifusing 1.5 ml microcentrifuge tubes, recover the DNA by centrifuga-tion at full speed for 1 min in a microcentrifuge.

7. Alternatively, if using 1.5 ml microcentrifuge tubes, recover theplasmid DNA by centrifugation at full speed for 1 minute ina microcentrifuge.

Note: The recovered elution volume will be 40–60% of the appliedvolume for the MicroPlex 24 spin method of sample recovery. DNArecovery will be approximately 30% to 40% lower than that with themicrofuge procedure.

● p18

Troubleshooting

Problem: The DNA yield is low.

Possible causes/solutions

1. The gel band was not fully dissolved: Incubate the gel band at 60 ˚Cfor the full 15 min. Visually inspect the sample to make sure it isdissolved before proceeding. Make sure that the size of the gel banddoes not exceed 300 mg and that the proper volume of capturebuffer is added.

2. The wash buffer was not completely removed before elution: Makesure that the GFX column is centrifuged for at least 30 s before theelution buffer is added. If humidity is high, increase the spin timeto 1 min.

Problem: The DNA sample floats out of the well when loading a gel.

Possible causes/solutions

1. The wash buffer was not completely removed before elution: Makesure that the GFX column is centrifuged for at least 30 s before theelution buffer is added. If humidity is high, increase the spin timeto 1 min.

2. Some of the final wash remained in the column below the frit andwas collected in the final elution: The collection tube was notemptied after the initial sample was spun through the column. Thiscaused the collection tube to overfill when the wash step was per-formed. Empty the collection tube as described in the procedure.If necessary, place the column back into the collection tube and spinbriefly to remove any residual fluid.

● p19

Problem: DNA does not cut to completion with restrictionenzymes/does not ligate.

Possible causes/solutions

1. Some of the final wash remained in the column below the frit andwas collected in the final elution: The collection tube was notemptied after the initial sample was spun through the column. Thiscaused the collection tube to overfill when the wash step was per-formed. Empty the collection tube as described in the procedure.If necessary, place the column back into the collection tube and spinbriefly to remove any residual fluid.

If problems persist, please contact GE Healthcare TechnicalService for assistance.

● p20

References1. Vogelstein, B. and Gillespie, D., Proc. Natl. Acad. Sci. USA 76, 615 (1979).

2. Marko, M. A., Chipperfield, R. and Birnboim, H. C., Anal. Biochem. 121, 382(1982).

● p21

Companion products available fromGE HealthcareProduct Product number

Taq DNA Polymerase See GE Healthcare Catalog

Ready-To-Go™ PCR Beads (100 reactions in 0.5 ml tubes) 27-9558-01

MicroPlex 24 See GE Healthcare Catalog

MicroPlex 24 Vacuum See GE Healthcare Catalog

MicroPlex 24 V-Bottom Collection Trays See GE Healthcare Catalog

MicroPlex 24 Vacuum Gaskets See GE Healthcare Catalog

MicroPlex 24 Vacuum Accessory Kit See GE Healthcare Catalog

GenomicPrep™ Blood DNA Isolation Kit 27-5236-01

GenomicPrep Cells and Tissue DNA Isolation Kit 27-5237-01

DNA Polymerization Mix 27-2094-01

50 Base-Pair Ladder 27-4005-01

100 Base-Pair Ladder 27-4007-01

KiloBase DNA Marker 27-4004-01

A full range of sequencing kits and reagents can be found in the GE Healthcare catalog.

● p22

All goods and services are sold subject to the terms and conditions of sale of the company within theGeneral Electric Company group which supplies them. A copy of these terms and conditions isavailable on request.

* U.S. Patent No. 5,603,899 has been issued to Pharmacia Biosciences for multiple column chromatography.

† The Polymerase Chain Reaction (PCR) is covered by patents owned by Roche Molecular Systems andF Hoffmann-La Roche Ltd. A license to use the PCR process for certain research and developmentactivities accompanies the purchase of certain reagents from licensed suppliers such as GE Healthcareand affiliates when used in conjuction with an authorized thermal cycler.

Printed in the United States.

● p23

● p24

GE Healthcare UK Ltd

GE Healthcare Place, Little Chalfont, Buckinghamshire, England HP7 9NA

tel 0870 606 1921 fax 01494 544350

GE Healthcare Bio-Sciences AB

SE-751 84 Uppsala, Sweden

tel 46 (0) 1816 5000 fax 46 (0) 1816 6458

GE Healthcare Bio-Sciences Corp

800 Centennial Avenue, PO Box 1327, Piscataway, NJ 08855 USA

tel 1-800-526-3593 fax 1-800-329-3593 or 877-295-8102

GE Healthcare Europe GmbH

Munzinger Strasse 9, D-79111, Freiberg, Germany

tel 0761 4903 01 fax 0761 4903 405

TrademarksAGE Healthcareare trademarks ofGeneral Electric Company.

GFX, MicroSpin, MicroPlex,Ready-To-Go andGenomicPrep aretrademarks of GE Healthcare Bio-Sciences Ltd.