GE Oven RPN2510E User Manual

GE Healthcare

Amersham

Hybridization oven/shaker

Product User Manual

Codes:

RPN2510E

RPN2511E

RPN2512E

1. Legal

GE, imagination at work and GE monogram are trademarks of General Electric

Company.

Amersham, Alkphos Direct, ECF, ECL, ECL Plus, ECL Advance, Hybond,

Hypercassette, Hyperfilm, Hyperscreen, Hypertorch, Megaprime, Nick, Rapid-

hyb, Ready-To-Go, Rediprime, Sensitive and Trackertape are trademarks of GE

Healthcare companies.

All third party trademarks are the property of their respective owners.

First published November 2003

All goods and services are sold subject to the terms and conditions of sale of the

company within GE Healthcare which supplies them.

A copy of these terms and conditions is available on request. Contact your local

GE Healthcare representative for the most current information.

http://www.gehealthcare.com/lifesciences

GE Healthcare UK Limited.

Amersham Place, Little Chalfont,

Buckinghamshire, HP7 9NA UK

RPN2510EUM Chapter 1 Rev C 06/2007

2. Introduction

One of the most widely used techniques in the molecular biology field is the

immobilization of DNA and RNA onto a solid support membrane and subsequent

hybridizations with a specific single stranded probe, labeled to facilitate its

detection.

Using the GE Healthcare Hybridization Oven/Shaker ensures that the temperature

and shaking/rotation frequency, and hence the stringency of hybridization and

washing steps are rigidly controlled. This enables rapid and reproducible probing of

nucleic acids, and proteins immobilized on nylon and nitrocellulose membranes.

The GE Healthcare Hybridization Oven/Shaker is a multipurpose instrument

combining accurate temperature control with a choice of interchangeable

hybridization modes:

• Variable speed rotisserie: holding 7 × 40 mm or 2 × 75 and 2 × 40 mm

hybridization bottles.

• A variable speed platform shaker for ‘box’ hybridizations, depurination,

denaturation and neutralization steps.

The instrument is:

• Economical: bottle hybridization minimizes probe volumes, reducing reagent

volumes and enhancing signal intensity.

• Precise: stringency of hybridization and washing steps are rigidly controlled,

ensuring reproducible results.

• Sensitive: validated protocols ensure optimal hybridization and washing steps,

enhancing multiple reprobing when using Hybond™ membranes.

• Safe: the double-glazed polycarbonate/acrylic door offers excellent thermal

insulation whilst minimizing radiation exposure.

• Convenient: small foot-print maximizes the use of limited laboratory space.

The instrument is suitable for use in conjunction with radiolabeled probes

using Rapid-hyb™ buffer, non-radioactive nucleic acid labeling and detection

systems such as AlkPhos Direct™, and protein labeling and detection systems

including ECL™, ECL Plus™ and ECL Advance™. Some protocols for use with these

applications are included in section 7.

RPN2510EUM Chapter 2 Rev C 06/2007

3. Safety warnings and

precautions

If the equipment is not used in the manner described in this manual the

protection provided might be impaired

This equipment is designed to operate under the following conditions: -

For indoor use only

Use in a well ventilated area

Ambient temperature range + 5°C to + 40°C

Altitude to 2000 m

Relative humidity not exceeding 80%

Mains supply fluctuation not exceeding 10%

Over-voltage category II IEC60364-4-443

Pollution degree 2

Use with a minimum distance all around of 200 mm from walls or other items

The unit should be carried using both hands.

Never move or carry the unit when in use or connected to the mains electricity supply.

In the case of mains interruption, a fault or electrical failure, the unit will continue

to operate on restoration of the electricity supply or removal of the fault.

3.1. Electrical Installation

WARNING: Lethal voltages inside the Hybridization Oven casing. Always

switch the Hybridization Oven/Shaker off and remove the plug from the

electrical socket before performing any cleaning or maintenance of the

instrument.

THIS INSTRUMENT MUST BE

EARTHED

IMPORTANT: The Hybridization Oven/Shaker has been supplied with a cordset and

plug. Check that the appropriate plug for your electrical supply has been included.

If there is any doubt consult a qualified electrician and inform your local GE

Healthcare sales office.

Should the plug require replacement in the future, the following information should

be noted:

The wires in the mains lead are coloured in accordance with the following code:

GREEN/YELLOW

BLUE

BROWN

Earth

Neutral

Live

As the colours of the wires in the mains lead of this item may not correspond with

the coloured markings identifying the terminals in your plug, proceed as follows:

• The wire which is coloured green/yellow must be connected to the terminal in

the plug which is marked with the letter ‘E’ or by the earth symbol or colours

green or green/yellow.

• The wire, which is coloured blue, must be connected to the terminal, which is

marked with the letter ‘N’ or coloured black.

• The wire, which is coloured brown, must be connected to the terminal, which is

marked ‘L’ or coloured red.

• Fit the plug with the appropriate fuse. Always replace the plug cover, never use

the plug without the cover.

The GE Healthcare Hybridization Oven/Shaker is intended for research use only

and not for any other purpose.

IF IN DOUBT CONSULT A QUALIFIED

ELECTRICIAN

RPN2510EUM Chapter 3 Rev C 06/2007

4. Specification

Overall dimensions

Height:

Depth:

Width:

17”

15”

(43.5 cm)

(38.0 cm)

Oven dimensions

Height:

Depth:

Width:

8”

9”

10”

(20.0 cm)

(23.0 cm)

(25.0 cm)

Weight:

24 kg

18 litres

Capacity (nominal):

Temperature range:

Temperature precision:

Temperature fluctuation:

Power rating:

Ambient plus 5°C–80°C

+/- 0.5°C

+/- 0.1°C (@37°C)

250 W

Over temperature cut-out:

Temperature variation:

Rotisserie speed:

1°C over set temperature

< 0.25°C

2–10 rev/minute

5–70 strokes/minute

Shaker platform speed:

Electrical

Nominal Voltage/Hertz/Amp

Product code

230 V / 50 Hz / 3.1 A

110/120 V / 60 Hz / 4 A

100 V / 50/60 Hz / 4 A

RPN2510E

RPN2511E

RPN2512E

Each instrument is supplied with 1 rotisserie (RPN2514),

6 hybridization bottles (RPN2516) and an instruction manual.

Accessories

Rotisserie, holds 7 × 40 mm hybridization bottles

RPN2514

RPN2515

RPN2516

RPN2517

RPN2518

RPN2519

Rotisserie, holds 2 × 75 mm and 2 × 40 mm hybridization bottles

Hybridization bottle, 260 × 40 mm (For rotisserie RPN2514, pack of 6)

Hybridization bottle, 170 × 40 mm (For rotisserie RPN2514, pack of 6)

Hybridization bottle, 240 × 75 mm (For rotisserie RPN2515, pack of 2)

Hybridization mesh, 1 roll 21.5 cm × 10 m

RPN2510EUM Chapter 4 Rev C 06/2007

5. Setting up the Hybridization

Oven/Shaker

1. Remove all packaging and place the Hybridization Oven/Shaker on a level

working surface, ensuring that there is sufficient room above the instrument to

allow the door to be opened fully.

2. Leave the unit to stand for a minimum of 3 hours. This will allow acclimation to

the new ambient temperature.

3. Plug the female end of the power cable into the Hybridization Oven/Shaker.

4. Connect the power cable to a suitably grounded electrical outlet. The correct

operating voltage of the Hybridization Oven/Shaker is found on the product

information label on the rear of the instrument.

5. Turn the Mains ON/OFF switch (1 on Fig 1) to the ON position.

6. The GE Healthcare Hybridization Oven/Shaker is now ready for use.

Rotisserie

Temperature (°C)

off

press

to set

off

rotisserie

(rev/minute)

shake r

(strokes/minute)

Hybridisation Oven/Shaker

Fig 1. Diagram of instrument control

panel

5.1. Setting the oven temperature

1. The Oven temperature controls are located on the right hand side of the control

panel (numbers 4 to 7 in figure 1).

Rotisserie/shaker controls

1. Rotisserie/shaker ON/OFF button

2. Rotisserie/shaker speed selector dial

3. Motor on indicator

2. To turn the temperature control ON press the on / off button (4). The LED display

(6) will show the current Oven temperature.

Temperature controls

4. Temperature on/off button

5. Press to set button

6. LED temperature display

7. Temperature selector dial

3. To set the temperature, press and hold the press to set button (5) and at

the same time rotate the temperature selector dial (7) until the required

temperature is shown on the digital display (6).

4. Release the press to set button (5) and the temperature display (6) will revert

back to the actual Oven temperature.

5. The set temperature can be viewed at any time simply by pressing and holding

the press to set button (6)

6. The oven will now automatically heat up to the set temperature.

Note: the Oven is fitted with automatic

digital over-temperature protection (see

Section 8.1. for more details).

5.2. Setting up the rotisserie

The rotisserie is installed in the Hybridization Oven/Shaker during transit. To use

the rotisserie for bottle hybridization, the following procedure should be adopted:

1. Lift up the oven door to its fullest extent to allow complete access to the oven

interior.

2. Lift the rotisserie vertically out of the oven. Unpack and place on the bench.

NOTE: Always ensure that the weight

is evenly distributed on both sides

of the rotisserie. Place an empty

hybridization bottle into the other side

of the rotisserie as a counterbalance if

necessary.

3. Place the membranes to be hybridized into the required number of hybridization

bottles. Using the rotisserie as a stand for the bottles, place the bottles into the

rotisserie, pushing them down as far as they will go.

RPN2510EUM Chapter 5 Rev C 06/2007

4. Place the rotisserie into the oven onto the rotation mechanism, ensuring that

the serrated bands at either end of the rotisserie locate onto the steel cogs

of the rotation mechanism at the rear of the oven. (B on figure 2). The plastic

flanges of the rotisserie locate on to the small wheels on the oven floor. Close

the oven door.

5. Ensure that the desired temperature has been set (see section 5.1).

6. The speed controls are on the left hand side of the control panel (numbers 1 to

3 in figure 1). Turn the Rotisserie/Shaker ON by pressing the on / off button (1).

The red indicator light (3) above the speed selector dial (2) will illuminate.

7. Turn the Rotisserie/Shaker speed selector dial (2) clockwise until the desired

rotation speed is reached (allowable values are 2–10 rpm).

The rotisserie will now start to rotate at the set speed.

8. When hybridization is complete turn the Rotisserie/Shaker OFF by pressing the

on / off button (1). The red indicator (3) will go out.

5.3. Setting up the platform shaker

During transit, the platform is stored vertically at the rear of the oven chamber. It

can remain in this position whilst the rotisserie is in use. To use the platform shaker

for ‘sandwich box’ hybridizations, the following procedure should be adopted:

1. Open the oven door to its fullest extent, lift the rotisserie vertically and store in a

safe place.

2. Lift the platform by its handle (A, see figure 2) from its storage position, slide it

forward and locate it on the rocking mechanism by placing the side pegs of the

platform into the retainers (D) on the side walls of the oven chamber. This action

seats the nylon blocks (C) on the underside of the platform on to the pegs (E)

protruding from the rocker mechanism at the rear of the oven.

3. Place the box in which the hybridization is being performed on to the shaker

platform and close the oven door.

4. Ensure that the desired temperature has been set (see section 4.1).

5. Turn the Rotisserie/Shaker ON by pressing the on / off button (1). The red

indicator light (3) above the speed selector dial (2) will illuminate.

6. Rotate the Rotisserie/Shaker speed selector dial (2) clockwise, until the desired

shaker speed is reached. Allowable values are 5–70 strokes per minute.

The shaker platform will now oscillate at the set speed.

7. When hybridization is complete turn the Rotisserie/Shaker OFF by pressing the

on / off button (1). The red indicator will go out (3).

Fig 2. Hybridization oven drive

components

RPN2510EUM Chapter 5 Rev C 06/2007

6. Hybridization using the platform

shaker

The Hybridization Oven/Shaker is compatible with the hybridization technologies

available from GE Healthcare. These include radioactive hybridizations using

Rapid hyb buffer and the range of non-radioactive systems for the labelling and

detection of proteins and nucleic acids (see appendix 1).

When using the platform shaker the hybridization and washing conditions

recommended in the appropriate GE Healthcare literature should

be used. The following protocol therefore provides a guideline. For specific

hybridization times and temperatures, refer to the relevant protocol booklet.

1. Prepare blots as recommended in the appropriate Hybond protocol booklet.

2. Set the oven temperature as described in section 5.1.

3. Place the membrane in a suitable box (or bag) and cover with sufficient

prehybridization buffer to ensure that the entire surface of the membrane is

covered. Recommended volume: surface area ratio is given in most protocol

booklets. Seal the box (or bag) securely.

4. Place the box (or bag) on the platform, set the oscillation speed to 30 strokes/

minute as described in section 5.3. Prehybridize for the required length of time.

5. Remove the box (or bag) from the oven and carefully add the labeled single

stranded probe (denaturation may be required post labelling refer to the

appropriate protocol booklet) to the prehybridization buffer.

NOTE: Do not pipette the probe solution

directly on to the membrane as this

may cause localized background.

6. Reseal the box (or bag) securely, replace it on the platform and hybridize for the

required length of time.

7. Remove the membranes and place in a clean box containing the first stringency

wash solution.

8. Increase the oscillation speed of the platform to 60 strokes/minute. Carry out

the recommended washing protocol at the appropriate temperature and for the

appropriate times.

RPN2510EUM Chapter 6 Rev C 06/2007

7. Hybridization using the rotisserie

Several advantages are associated with performing hybridizations in bottles,

namely those of safety and economy as outlined in the introduction. However,

the use of bottles for hybridizations and washing procedures requires certain

adaptations to standard protocols.

7.1. Assembly of membranes into bottles

1. Add approximately 20 ml 2 × SSC buffer into the hybridization bottle. The

rotisserie acts as a convenient bottle stand.

2. Pre-wet the membrane in 2 × SSC buffer and loosely roll it up.

3. Insert the rolled up membrane into the bottle and replace the cap. Ensure

that the cap is screwed on securely, (hand tight plus a quarter turn). DO NOT

OVERTIGHTEN, or the thread of the cap can be damaged, leading to leakages.

NOTE: If placing several small blots into

one bottle, prewet the membranes as

above, and space them out along the

length of the bottle with forceps.

4. Place the bottle on a flat surface and roll it gently in the opposite direction to

that in which the membrane is coiled. This rolling action causes the membrane

to uncoil, so lining the inner surface of the bottle.

NOTE: The use of a mesh in bottle

hybridizations to ensure uniform

contact between the membrane

surface and the buffer has been

recommended.

However, studies at GE Healthcare laboratories using a wide range of hybridization

mesh technologies demonstrate a resulting loss of sensitivity due to partial

absorption of the probe into the mesh.

A nylon mesh (RPN2519) is available as an optional extra, as it can facilitate easier

handling of a number of blots and the more fragile nitrocellulose membranes.

These handling advantages should be considered against the potential loss of

sensitivity before use.

When a hybridization mesh is used in conjunction with the membrane, the

following procedure should be adopted:

5. Pre-wet the mesh alongside the membrane in 2 × SSC buffer and place the

prewetted membrane on top of the mesh. The mesh should be slightly larger

than the blot in all dimensions. Roll both up together, with the mesh on the

outside of the membrane, and insert into the bottle as described above (7.1.4.).

7.2. Hybridization

1. Set the required oven temperature as detailed in section 5.1.

2. Discard the 2 × SSC, used in the bottle to unroll the membrane, and replace

with prehybridization buffer. Recommended volumes are provided in most GE

Healthcare protocol booklets. Generally a minimum of 10–15 ml per 20 × 20 cm

blot is advised. Seal the bottle, avoiding overtightening.

3. Place the hybridization bottle(s) into the rotisserie (as detailed in section 5.2.) add

counterbalance bottles if necessary. Place the rotisserie into the oven so that

the bottles are rotating in the same direction as the membrane is rolled, see

Figure 3.

Fig 3. Rotation of rotisserie

rotation of

membrane

RPN2510EUM Chapter 7 Rev C 06/2007

4. Prehybridize the membrane for the specified length of time at a rotisserie speed

of 4 rpm.

5. Following prehybridization, turn off the rotisserie, remove the rotisserie from the

oven. Add the labeled probe to the buffer, either by removing an aliquot of the

buffer, adding the probe and returning the aliquot to the bottle; or by adding the

probe directly into the bottle, avoiding the membrane.

6. Hybridize for the specified length of time at a rotisserie speed of 4 rpm.

7.3. Membrane washing procedures

Membranes can either be removed from the hybridization bottles and washed in a

box on the platform shaker (this is a more efficient procedure), or the washing

procedure may be carried out in the bottles. If the platform shaker is used, follow

the standard washing procedure mentioned in the appropriate protocol booklet.

If bottles are to be used it is necessary to modify the standard washing procedure.

Outlined in this section are the optimized washing protocols for radioactive

hybridizations and non-radioactive based hybridizations.

7.3.1. Radioactive Hybridizations

1. Carefully drain off the hybridization buffer, rinse the bottle and the membrane

thoroughly with 2 × SSC and discard.

NOTE: This last step is a high stringency

wash and should be omitted if related

sequences are to be probed.

2. Perform the following stringency washes in large volumes (100 ml minimum) of

the following solutions, at a rotisserie speed of 8 rpm:

2 × 10 minutes with 2 × SSC, 0.1% SDS at 65°C

1 × 15 minutes with 1 × SSC, 0.1% SDS at 65°C

2 × 10 minutes with 0.1 × SSC, 0.1% SDS at 65°C

(More washes over the same time period for each stringency condition can

improve background).

3. Remove the membrane from the bottle, drain off excess stringency wash, wrap

in SaranWrap™ and autoradiograph.

7.3.2. AlkPhos Direct Hybridizations

4. Drain off the hybridization buffer, rinse the bottle and the membrane thoroughly

with primary wash buffer and discard.

5. Perform the following stringency washes in large volumes (100 ml minimum) of

the following solutions, at a rotisserie speed of 8 rpm:

3 × 10 minutes with primary wash buffer solution at 55°C

NOTE: The room temperature washes

can be achieved by switching off

the oven and leaving the door open

whilst performing the washes or by

allowing the oven to cool down to room

temperature before performing the

final washes.

3 × 5 minutes with secondary wash buffer at room temperature

6. Remove the membrane from the bottle and detect using the standard

procedures outlined in the protocol booklet.

RPN2510EUM Chapter 7 Rev C 06/2007

8. Maintenance/care/cleaning of

the oven/shaker

The GE Healthcare Hybridization Oven/Shaker is designed to provide trouble-free

operation. The base of the oven and the shaker tray act as a spills tray and will

contain any spillage that occurs during hybridization and washing procedures.

To ensure lasting operation the following instructions should be followed:

ALWAYS DISCONNECT THE HYBRIDIZATION OVEN/SHAKER FROM THE

ELECTRICAL SUPPLY BEFORE CLEANING OR DRYING THE INSTRUMENT.

1. Any leakage from the hybridization bottles or the sandwich boxes should be

cleaned up immediately. Do not allow any liquids to enter the drive mechanism.

2. Wipe away any liquids from inside and outside of the unit using soap and water

with a soft cloth or sponge.

3. Do not allow chemicals to remain on unit surfaces.

4. Never clean unit with abrasive pads or cleaners.

5. Never clean unit with acetone or chloroform.

Only spare parts supplied or specified by GE Healthcare or its agent should

be used. Fitting of non-approved parts may affect the performance of the

safety features designed into the instrument.

8.1. Error codes/faults

The Oven has built in fault diagnostics. If a fault occurs, this system will display

an error code in the LED display to help the service engineer rectify the problem.

Please see the table below for details of error codes and other faults. In the

unlikely event that a problem does occur, note down which code / fault you

observe and contact your nearest GE Healthcare Office for further assistance. If

you have your own service personnel available for repair, a comprehensive service

manual is available on request.

Fault

Safety system in operation

Fault displayed by

Temperature probe PT100 reading low

Automatic over-temp system activated

LED displays Err and 0.1 alternatively

LED displays Err and 0.2 alternatively

Top left dot flashes in display

Temperature probe PT100 reading high Automatic over-temp system activated

Triac fault

Secondary relay control activated

Cuts power to motor

Rotisserie / shaker motor stalled

No movement seen but Rotisserie red

indicator light still on

Software crash

Cuts mains power

No temperature or motor motor power,

all LED’s blank

RPN2510EUM Chapter 8 Rev C 06/2007

9. Troubleshooting guide

This section briefly summarizes some of the potential problems encountered during membrane hybridizations. More

complete troubleshooting guides are supplied in the pack leaflet that accompany each product.

Symptom

Cause

Remedy

1. Membrane curling up in

hybridization bottle

1.1. Incorrect orientation of bottle in

1.1. Ensure membrane is rolled up in

the same direction as the bottle is

rotating (see section 7.2)

rotisserie

2. High background

2.1. Probe concentration too high

2.1. Reduce probe concentration

2.2. Unincorporated P nucleotides not 2.2. Remove unincorporated

removed

nucleotides

2.3. Insufficient blocking

2.3. Use recommended hybridization

buffer or extend prehybridization

time

2.4. Insufficient washing

2.4. Increase number of buffer changes

during the washing stage or

increase stringency of final wash

3. Weak signal

3.1. No transfer from gel to membrane

3.1. Load extra lanes with control DNA.

Transfer can be checked by

restaining gel with ethidium

bromide. If large DNA fragments

are detected poorly, use a

depurination step (0.25 M HCI)

3.2. Probe not labeled

3.2. Check probe labelling before

hybridization. See protocol in probe

labelling booklet

3.3. Probe not denatured

3.3. Boil probe for 5 minutes before

adding to hybridization buffer

3.4. Low specific activity of probe

3.5. Washes too stringent

3.4. Review labelling conditions

3.5. Increase buffer salt concentration

and decrease temperature

4. Patchy backgrounds

4.1. Hybridization buffer or wash

solution not evenly covering

membrane

4.1. Increase volume of hybridization

wash or solutions or use mesh (see

section 7.1. step 4)

4.2. Damaged membrane

4.2. Handle membrane carefully with

forceps

5. High background around edge of

membrane

5.1. Damaged membrane

5.1. Use clean scissors or a sharp

scalpel to cut membrane

RPN2510EUM Chapter 9 Rev C 06/2007

Appendix 1. Products for

electrophoresis

DNA Markers

Precise Sizing

Digest of Natural DNAs

50 Base-pair 100 Base-pair

KiloBase

DNA marker

27-4004-01

DRigest III

27-4060-01

ladder

27-4005-01

27-4007-01

500^a

450

400

350

300

250

200

150

100

2000^a

1900

1800

1700

1600

1500

1400

1300

1200

1100

1000

900

10000

8000

6000

5000

4000

3000

2500

2000

1500

1000

500

23130^b

9416

6557

4361^b

2322

2027

1353

1078

872

603

564^d

310

800

281

700

271

600

234

500

194

400

125^c

118

300

200

100

a not necessarily the largest size fragment possible, only the largest that is

readily distinguishable

b cos ends are located on these bands

c may not always be visible

Marker code number

27-4005-01

Recommended gel

2% agarose/6% polyacrylamide

1.5% agarose

Loading amount (µg/lane)

Heat before loading

2.0

27-4007-01

27-4004-01

0.8% agarose

0.5

27-4060-01

1% agarose

0.5–1.0

60–65°C for 2 minutes

RPN2510EUM Appendix 1 Rev C 06/2007

Appendix 2. Hybond membranes

for nucleic acid applications

Hybond ECL Hybond C Extra Hybond P

Size

Pack size

Hybond N+

Hybond XL

Hybond N

Hybond NX

82 mm

50 discs

RPN82B

RPN87B

RPN82S

RPN87S

RPN132S

RPN137S

RPN82N

RPN87N

RPN132N

RPN137N

RPN82T

RPN87T

RPN82D

RPN82E

87 mm

132 mm

50 discs

RPN132B

RPN137B

RPN1782B

RPN1787B

RPN1732B

RPN1737B

RPN3050B

RPN132T

RPN137T

RPN132D

137 mm

50 discs

RPN137E

82 mm

50 gridded discs

5 sheets

87 mm

132 mm

137 mm

30 × 50 cm

9 × 10 cm

10 × 10 cm

15 × 20 cm

20 × 20 cm

22.2 × 22.2 cm

14 × 16 cm

12 × 10 cm

15 × 10 cm

6 × 8 cm

RPN3050S

RPN3050N

10 sheets

15 sheets

20 sheets

50 sheets

1 roll

RPN910D

RPN1010D

RPN1520D

RPN2020D

RPN1520B

RPN2020B

RPN2222B

RPN1520S

RPN2020S

RPN2222S

RPN1520N

RPN2020N

RPN2222N

RPN1520T

RPN2002T

RPN2020E

RPN2020F

RPN1416F

RPN1210B

RPN1510B

RPN1210S

RPN1510S

RPN1210N

RPN1510N

RPN68D

RPN78D

7 × 8 cm

11.9 × 7.8 cm

22.2 × 22.2 cm

22.5 × 22.5 cm

115 × 73 mm

20 cm × 3 m

30 cm × 3 m

RPN119B

RPN2250B

RPN225B

RPN1576B

RPN203B

RPN303B

RPN119S

RPN119N

RPN203S

RPN303S

RPN203N

RPN303N

RPN203D

RPN303D

RPN3032D

RPN203E

RPN303E

1 roll

RPN303T

RPN303F

1 roll

a = 0.2 µm pack size

RPN2510EUM Appendix 2 Rev C 06/2007

Appendix 3. Radioactive labeling

systems

Labelling

system

Technology

Nucleotide

Amount of

template

Labelling

time

Probe specific

activity

Recommended

application

Product

code

(dpm/µg)

Rediprime

™

Random-prime

dCTP only

25 ng

10 minutes

5 minutes

10⁹

membrane

hybridization

RPN1633

RPN1634

Ready-To-Go

DNA labelling

beads

™

10 ng–1 µg

membrane

hybridization

DNA labelling

27-9240-01

Megaprime

™

Random-prime

Nick translation

any dNTP

25 ng

1 µg

10 minutes

2–3 hours

10⁹

membrane

hybridization

RPN1604

RPN1605

RPN1606

RPN1607

Nick translation

production of

large amounts

of probe

N5000

N5500

RPN2510EUM Appendix 3 Rev C 06/2007

Appendix 4. Non-radioactive

labeling and detection systems

Labelling and detection

system

Sensitivity

Time from

hybridization light output

to detection

Duration of

Strip before Quantification Recommended application

re-probing

AlkPhos Direct (RPN3690)

ECL Direct (RPN3000)

0.06 pg

0.5 pg

1 hour

5 days

yes

Single copy Southern and

Northerns

1 hour

1–2 hours

High target applications

eg. colony/plaques

RPN2510EUM Appendix 4 Rev C 06/2007

Appendix 5. Products for

autoradiography and

chemiluminscent detection

Hyperfilm – high performance autoradiography films

Product description

size

number sheets

code

Hyperfilm ECL

Hyperfilm MP

Hyperfilm MP Enveloped

7 inches

100

28-9068-35

28-9068-36

28-9068-37

28-9068-38

28-9068-39

28-9068-40

28-9068-41

28-9068-42

28-9068-43

28-9068-44

28-9068-45

28-9068-46

28-9068-47

28-9068-48

28-9068-50

24 cm

10 inches

30 cm

43 cm

7 inches

24 cm

10 inches

30 cm

43 cm

24 cm

Hypercassette™ – cassettes for autoradiography and light detection

Size

Code (standard)

Code (deep)

24 cm

30 cm

40 cm

43 cm

40 cm

RPN11642

RPN11643

RPN11644

RPN11645

RPN11646

RPN11647

RPN11648

RPN11649

RPN11650

RPN11628

RPN11627

7 inches

10 inches

RPN11629

10 12 inches

125

Hyperscreen™ – intensifying screens for P and

I autoradiography

Size

Code

Quantity

24 cm

30 cm

40 cm

43 cm

40 cm

RPN1662

RPN1663

RPN1664

RPN1665

RPN1666

RPN1667

RPN1668

RPN1669

RPN1670

1 pair

7 inches

10 inches

10 12 inches