GE RPN2512E, RPN2510E, RPN2511E User Manual

GE Healthcare

Amersham Hybridization oven/shaker

Product User Manual

Codes: RPN2510E

RPN2511E RPN2512E

Page finder

1. Legal 3

2. Introduction 4

3. Safety warnings and precautions 5

3.1. Electrical Installation 5

4. Specification 6

5. Setting up the Hybridization Oven/Shaker 7

5.1. Setting the oven temperature 7

5.2. Setting up the rotisserie 7

5.3. Setting up the platform shaker 8

6. Hybridization using the platform shaker 9

7. Hybridization using the rotisserie 10

7.1. Assembly of membranes into bottles 10

7.2. Hybridization 10

7.3. Membrane washing procedures 11

7.3.1. Radioactive Hybridizations 11

7.3.2. AlkPhos Direct Hybridizations 11

8. Maintenance/care/cleaning of the oven/shaker 12

8.1. Error codes/faults 12

9. Troubleshooting guide 13

Appendix 1. Products for electrophoresis 14

Appendix 2. Hybond membranes for nucleic acid applications 15

Appendix 3. Radioactive labeling systems 16

Appendix 4. Non-radioactive labeling and detection systems 17

Appendix 5. Products for autoradiography and chemiluminscent

detection 18

Appendix 6. Hybridization buffer 19

Appendix 7. Radiation safety products 20

RPN2510EUM Pagefinder Rev C 06/2007 2

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, Rapidhyb, Ready-To-Go, Rediprime, Sensitive and Trackertape are trademarks of GE Healthcare companies.

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

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

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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.

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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.

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 Earth BLUE Neutral BROWN 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.

THIS INSTRUMENT MUST BE EARTHED

• 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.

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IF IN DOUBT CONSULT A QUALIFIED ELECTRICIAN

4. Specification

Overall dimensions

Height: 17” (43.5 cm) Depth: 15” (38.0 cm) Width: 15” (38.0 cm)

Oven dimensions

Height: 8” (20.0 cm) Depth: 9” (23.0 cm) Width: 10” (25.0 cm)

Weight: 24 kg

Capacity (nominal): 18 litres

Temperature range: Ambient plus 5°C–80°C

Temperature precision: +/- 0.5°C

Temperature fluctuation: +/- 0.1°C (@37°C)

Power rating: 250 W

Over temperature cut-out: 1°C over set temperature

Temperature variation: < 0.25°C

Rotisserie speed: 2–10 rev/minute

Shaker platform speed: 5–70 strokes/minute

Electrical

Nominal Voltage/Hertz/Amp Product code

230V/50Hz/3.1A  RPN2510E 110/120V/60Hz/4A  RPN2511E 100V/50/60Hz/4A  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

Rotisserie, holds 2 × 75 mm and 2 × 40 mm hybridization bottles RPN2515

Hybridization bottle, 260 × 40 mm (For rotisserie RPN2514, pack of 6) RPN2516

Hybridization bottle, 170 × 40 mm (For rotisserie RPN2514, pack of 6) RPN2517

Hybridization bottle, 240 × 75 mm (For rotisserie RPN2515, pack of 2) RPN2518

Hybridization mesh, 1 roll 21.5 cm × 10 m RPN2519

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5. Setting up the Hybridization

Hybridisation Oven/Shaker

press

to set

Rotisserie

rotisserie (rev/minute)

shake r

(strokes/minute)

Te mperatur e (

°C )

on off

off

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.

1 2 3 4 5 6 7

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).

2. To turn the temperature control ON press the on / off button (4). The LED display (6) will show the current Oven temperature.

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.

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.

Fig 1. Diagram of instrument control panel

Rotisserie/shaker controls

1. Rotisserie/shaker ON/OFF button

2. Rotisserie/shaker speed selector dial

3. Motor on indicator

Temperature controls

4. Temperature on/off button

5. Press to set button

6. LED temperature display

7. Temperature selector dial

Note: the Oven is fitted with automatic digital over-temperature protection (see Section 8.1. for more details).

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

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.

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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.

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