GE RPN2512E, RPN2510E, RPN2511E User Manual

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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

RPN2510EUM Chapter 1 Rev C 06/2007 3

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 4

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.

RPN2510EUM Chapter 3 Rev C 06/2007 5

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

RPN2510EUM Chapter 4 Rev C 06/2007 6

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.

RPN2510EUM Chapter 5 Rev C 06/2007 7

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.

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).

CB A

Fig 2. Hybridization oven drive components

RPN2510EUM Chapter 5 Rev C 06/2007 8

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.

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.

NOTE: Do not pipette the probe solution directly on to the membrane as this may cause localized background.

RPN2510EUM Chapter 6 Rev C 06/2007 9

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.

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.

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.).

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.

NOTE: The use of a mesh in bottle hybridizations to ensure uniform contact between the membrane surface and the buffer has been recommended.

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.

rotation of membrane

Fig 3. Rotation of rotisserie

RPN2510EUM Chapter 7 Rev C 06/2007 10

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.

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 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.

NOTE: This last step is a high stringency wash and should be omitted if related sequences are to be probed.

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.

RPN2510EUM Chapter 7 Rev C 06/2007 11

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

Temperature probe PT100 reading high Automatic over-temp system activated LED displays Err and 0.2 alternatively

Triac fault Secondary relay control activated Top left dot flashes in display

Rotisserie / shaker motor stalled Cuts power to motor 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 12

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

1. Membrane curling up in

hybridization bottle

2. High background

3. Weak signal

Cause

1.1. Incorrect orientation of bottle in

rotisserie

2.1. Probe concentration too high

2.2. Unincorporated 32P nucleotides not

removed

2.3. Insufficient blocking

2.4. Insufficient washing

3.1. No transfer from gel to membrane

3.2. Probe not labeled

3.3. Probe not denatured

Remedy

1.1. Ensure membrane is rolled up in

the same direction as the bottle is rotating (see section 7.2)

2.1. Reduce probe concentration

2.2. Remove unincorporated

nucleotides

2.3. Use recommended hybridization buffer or extend prehybridization time

2.4. Increase number of buffer changes during the washing stage or increase stringency of final wash

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. Check probe labelling before hybridization. See protocol in probe labelling booklet

3.3. Boil probe for 5 minutes before adding to hybridization buffer

4. Patchy backgrounds

5. High background around edge of membrane

3.4. Low specific activity of probe

3.5. Washes too stringent

4.1. Hybridization buffer or wash

solution not evenly covering membrane

4.2. Damaged membrane

5.1. Damaged membrane

3.4. Review labelling conditions

3.5. Increase buffer salt concentration

and decrease temperature

4.1. Increase volume of hybridization wash or solutions or use mesh (see section 7.1. step 4)

4.2. Handle membrane carefully with forceps

5.1. Use clean scissors or a sharp scalpel to cut membrane

RPN2510EUM Chapter 9 Rev C 06/2007 13

Appendix 1. Products for electrophoresis

DNA Markers

Precise Sizing Digest of Natural DNAs

50 Base-pair 100 Base-pair KiloBase DRigest III ladder ladder DNA marker 27-4060-01 27-4005-01 27-4007-01 27-4004-01

500a 2000a 10000 23130

450 1900 8000 9416

400 1800 6000 6557

350 1700 5000 4361b

300 1600 4000 2322

250 1500 3000 2027

200 1400 2500 1353

150 1300 2000 1078

100 1200 1500 872

50 1100 1000 603

1000 500 564

900 310

800 281

700 271

600 234

500 194

400 125

300 118

200 72

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 Recommended gel Loading amount (µg/lane) Heat before loading

27-4005-01 2% agarose/6% polyacrylamide 2.0 No

27-4007-01 1.5% agarose 2.0 No

27-4004-01 0.8% agarose 0.5 No

27-4060-01 1% agarose 0.5–1.0 60–65°C for 2 minutes

RPN2510EUM Appendix 1 Rev C 06/2007 14

Appendix 2. Hybond membranes for nucleic acid applications

Size Pack size Hybond N+ Hybond XL Hybond N Hybond NX Hybond ECL Hybond C Extra Hybond P

82 mm 50 discs RPN82B RPN82S RPN82N RPN82T RPN82D RPN82E 87 mm 50 discs RPN87B RPN87S RPN87N RPN87T 132 mm 50 discs RPN132B RPN132S RPN132N RPN132T RPN132D 137 mm 50 discs RPN137B RPN137S RPN137N RPN137T RPN137E 82 mm 50 gridded discs RPN1782B 87 mm 50 gridded discs RPN1787B 132 mm 50 gridded discs RPN1732B 137 mm 50 gridded discs RPN1737B 30 × 50 cm 5 sheets RPN3050B RPN3050S RPN3050N 9 × 10 cm 10 sheets RPN910D 10 × 10 cm 10 sheets RPN1010D 15 × 20 cm 10 sheets RPN1520B RPN1520S RPN1520N RPN1520T RPN1520D 20 × 20 cm 10 sheets RPN2020B RPN2020S RPN2020N RPN2002T RPN2020D RPN2020E RPN2020F

22.2 × 22.2 cm 10 sheets RPN2222B RPN2222S RPN2222N 14 × 16 cm 15 sheets RPN1416F 12 × 10 cm 20 sheets RPN1210B RPN1210S RPN1210N 15 × 10 cm 20 sheets RPN1510B RPN1510S RPN1510N 6 × 8 cm 50 sheets RPN68D 7 × 8 cm 50 sheets RPN78D

11.9 × 7.8 cm 50 sheets RPN119B RPN119S RPN119N

22.2 × 22.2 cm 50 sheets RPN2250B

22.5 × 22.5 cm 50 sheets RPN225B 115 × 73 mm 50 sheets RPN1576B 20 cm × 3 m 1 roll RPN203B RPN203S RPN203N RPN203D RPN203E 30 cm × 3 m 1 roll RPN303B RPN303S RPN303N RPN303T RPN303D RPN303E RPN303F 30 cm × 3 m 1 rolla RPN3032D

a = 0.2 µm pack size

RPN2510EUM Appendix 2 Rev C 06/2007 15

Appendix 3. Radioactive labeling systems

Labelling Technology Nucleotide Amount of Labelling Probe specific Recommended Product system template time activity application code (dpm/µg)

Rediprime™ II Random-prime dCTP only 25 ng 10 minutes 2 × 109 membrane RPN1633

hybridization RPN1634

Ready-To-Go™ Random-prime dCTP only 10 ng–1 µg 5 minutes 2 × 109 membrane 27-9240-01 DNA labelling hybridization beads DNA labelling

Megaprime™ Random-prime any dNTP 25 ng 10 minutes 2 × 109 membrane RPN1604

hybridization RPN1605 RPN1606 RPN1607

Nick translation Nick translation any dNTP 1 µg 2–3 hours 2 × 109 production of N5000 large amounts N5500 of probe

RPN2510EUM Appendix 3 Rev C 06/2007 16

Appendix 4. Non-radioactive labeling and detection systems

Labelling and detection Sensitivity Time from Duration of Strip before Quantification Recommended application system hybridization light output re-probing to detection

AlkPhos Direct (RPN3690) 0.06 pg 1 hour 5 days yes no Single copy Southern and

Northerns

ECL Direct (RPN3000) 0.5 pg 1 hour 1–2 hours no no High target applications eg. colony/plaques

RPN2510EUM Appendix 4 Rev C 06/2007 17

Appendix 5. Products for autoradiography and chemiluminscent detection

Hyperfilm – high performance autoradiography films

Product description size number sheets code

Hyperfilm ECL 5 × 7 inches 50 28-9068-35 Hyperfilm ECL 18 × 24 cm 50 28-9068-36 Hyperfilm ECL 18 × 24 cm 100 28-9068-37 Hyperfilm ECL 8 × 10 inches 50 28-9068-38 Hyperfilm ECL 8 × 10 inches 100 28-9068-39 Hyperfilm ECL 24 × 30 cm 50 28-9068-40 Hyperfilm ECL 35 × 43 cm 50 28-9068-41 Hyperfilm MP 5 × 7 inches 50 28-9068-42 Hyperfilm MP 18 × 24 cm 50 28-9068-43 Hyperfilm MP 18 × 24 cm 100 28-9068-44 Hyperfilm MP 8 × 10 inches 50 28-9068-45 Hyperfilm MP 8 × 10 inches 100 28-9068-46 Hyperfilm MP 24 × 30 cm 50 28-9068-47 Hyperfilm MP 35 × 43 cm 50 28-9068-48 Hyperfilm MP Enveloped 18 × 24 cm 50 28-9068-50

Hypercassette™ – cassettes for autoradiography and light detection

Size Code (standard) Code (deep)

18 × 24 cm RPN11642 RPN11628 24 × 30 cm RPN11643 30 × 40 cm RPN11644 RPN11627 35 × 43 cm RPN11645 18 × 43 cm RPN11646 20 × 40 cm RPN11647 5 × 7 inches RPN11648 8 × 10 inches RPN11649 RPN11629 10 × 12 inches RPN11650

Hyperscreen™ – intensifying screens for 32P and

Size Code Quantity

18 × 24 cm RPN1662 1 pair 24 × 30 cm RPN1663 1 pair 30 × 40 cm RPN1664 1 pair 35 × 43 cm RPN1665 1 pair 18 × 43 cm RPN1666 1 pair 20 × 40 cm RPN1667 1 pair 5 × 7 inches RPN1668 1 pair 8 × 10 inches RPN1669 1 pair 10 × 12 inches RPN1670 1 pair

125

I autoradiography

GE RPN2512E, RPN2510E, RPN2511E User Manual

Specifications and Main Features

Frequently Asked Questions

User Manual