Thermo Scientific Shake n Stack 6240 Operating instructions

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User Manual
Shake ‘n Stack
Hybridisation Oven
Operating and Maintenance Manual 7026240 Rev. 1

Thermo Scientific
Manual Number 7026240
1 ECNM-000224 02/06/2020 Removed model numbers 6244 & 6245 from the manual. TFI
0 25036/OV-368 8/16/11 New controller - Release 3 ccs
EV EC /ECN DATE DESC IPTION By
Preface
Shake ‘n Stack i
Models covered by this manual
Models Voltage Includes
6240 220 Shaking platform, drip tray, rotisserie
6241 110 Shaking platform, drip tray, rotisserie
6242 220 Drip tray, rotisserie
6243 110 Drip tray, rotisserie

Thermo Scientificii Shake ‘n Stack
Preface
Contains Parts and Assemblies
Susceptible to Damage by
Electrostatic Discharge (ESD)
CAUTION
Important Read this instruction manual. Failure to read, understand and follow the instructions in this manual
may result in damage to the unit, injury to operating personnel, and poor equipment performance. s
Caution All internal adjustments and maintenance must be performed by qualified service personnel. s
Material in this manual is for information purposes only. The contents and the product it describes are subject
to change without notice. Thermo Fisher Scientific makes no representations or warranties with respect to this
manual. In no event shall Thermo be held liable for any damages, direct or incidental, arising out of or related to
the use of this manual.
© 2020 Thermo Fisher Scientific. All rights reserved.

Thermo Scientific Shake ‘n Stack iii
Preface
Important operating and/or maintenance instructions. Read the accompanying text carefully.
Potential electrical hazards. Only qualified persons should perform procedures associated with this
symbol.
Equipment being maintained or serviced must be turned off and locked off to prevent possible injury.
ot surface(s) present which may cause burns to unprotected skin, or to materials which may be
damaged by elevated temperatures.
Marking of electrical and electronic equipment, which applies to electrical and electronic equipment
falling under the Directive 2012/19/EU (WEEE) and the equipment that has been put on the market
after 13 August 2005.
This product is required to comply with the European Union’s Waste Electrical & Electronic
Equipment (WEEE) Directive 2012/19/EU. It is marked with the WEEE symbol. Thermo Fisher
Scientific has contracted with one or more recycling/disposal companies in each EU Member State
European Country, and this product should be disposed of or recycled through them. Further
information on Thermo’s compliance with this directive, the recyclers in your country and
information on Thermo Scientific products will be available at www.thermofisher.com.
4Always use the proper protective equipment (clothing, gloves, goggles, etc.)
4Always dissipate extreme cold or heat and wear protective clothing.
4Always follow good hygiene practices.
4Each individual is responsible for his or her own safety.

Thermo Scientificiv Shake ‘n Stack
Preface
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Thermo Scientific
Table of Contents
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-1
Oven Temperature Monitoring . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-1
Safety Precautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-2
Unpacking and Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-1
Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-2
Set the Required Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .2-2
Set Up the Shaking Platform . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3-1
Methodology of Hybridisation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-1
Place Membranes in a Bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-1
Insert Membranes in a Bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-2
Pre- ybridisation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-3
ybridisation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-3
Washing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-4
Washing in ybridisation Bottles . . . . . . . . . . . . . . . . . . . . . . . . . .4-4
Washing in a Sandwich Box . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4-4
otisseries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-1
Variable Axis Rotisseries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-1
50ml & 15ml Tube Rotisseries . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5-2
Section 1
Section 2
Section 3
Section 4
Section 5
Shake ‘n Stack v

vi Shake ‘n Stack Thermo Scientific
Cleaning and Decontamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-1
Meshes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-1
Bottles & Caps . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-2
Oven Interior/Shaking Platform . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-2
Rotisserie . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .6-2
Use of adioactive Probes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7-1
Technical Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8-1
Additional Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8-3
Exploded Parts Drawings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8-4
Electrical Schematics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9-1
Warranty Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10-1
Troubleshooting Guide for Nucleic Acid Hybridisations . . . . . . . . .AI-1
Background Reduction - General . . . . . . . . . . . . . . . . . . . . . . . . . . .AI-1
Factors Resulting in igh Backgrounds . . . . . . . . . . . . . . . . . . . . . .AI-3
Factors Resulting in Signal Lower than Expected . . . . . . . . . . . . . . .AI-4
Hybidisation Bottle Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .AII-1
Bottle Care . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .AII-1
Table of Contents
Section 6
Section 8
Section 7
Section 9
Section 10
Appendix I
Appendix II

Shake ‘n Stack 1-1Thermo Scientific
Oven Temperature
Monitoring
Section 1 Introduction
The Thermo system of ybridising in bottles is considered by many
molecular biologists to be the best method for conducting ybridisations
with Southern, Northern, Dot, Slot or Colony Blots. Additional
information, including detailed protocols, is contained in the included
ybridisation Guide.
ybridising in bottles means that probe volumes may be significantly
reduced compared to experiments performed in conventional systems, and
the continual movement of the probe across the surface of the membrane
results in very efficient hybridisation reactions.
Protection from exposure to radiation is provided by the heavy walled
borosilicate glass bottles.
Warning Based on tests, these bottles block almost 100% of BETA
radiation emissions of a 1,000 pCi P-32 source. owever, users should
observe precautions due to induced X-ray phenomenon (Breemsstrahlung)
occurring, depending on the type(s) of radionuclides used. s
The ybridisation Oven itself provides additional shielding. In the event
of a spillage within the Oven, the stainless steel drip tray will contain up to
6.8 fl. oz. (200 ml) of liquid.
Note If the Shake ‘n Stack is not used as specified in this manual, the
protection provided by the equipment may be impaired.
All Ovens are calibrated by matching the thermistor temperature read-out,
seen on the LCD display, to the actual measurement of distilled water
contained within a ybridisation Bottle as it rotates. The temperature
measuring device used inside the calibration bottle is traceable to national
standards.
This calibration method ensures all experiments using the rotisserie meet
our temperature accuracy specification of ±1°C and indeed, due to the
rotisserie action, uniformity within the bottle is ±0.25°C. owever, it is
important to note that the temperature inside the oven chamber will vary
from point to point and therefore, the temperature of objects/vessels,
which are not placed in the rotisserie, will vary from that which is
displayed. Refer to “Setting the Required Temperature” for
recommendations on temperature setting when using shaking platforms.

1-2 Shake ‘n Stack Thermo Scientific
Section 1
Introduction
The Shake ‘n Stack is designed to be used with Thermo’s hybridisation
bottle. These high quality leak-proof containers can be used at
temperatures up to 70°C (for catalogue numbers, see Section 8).
Bottles should not be used above 70°C without taking the precaution of
releasing the pressure formed by the release of gases from solutions as they
are warmed. Simply loosening, then retightening the bottle cap at
approximately 60°C should relieve the pressure.
Caution Dropping or over-tightening lids can cause cracking and failure.
Lids should be inspected prior to each use. s
Caution Always wear gloves when handling the bottle and use the bottle
gripper provided (see Appendix II for full instructions on Bottle Care). s
Finally, the Shake ‘n Stack Oven is designed for reliability and ease of
maintenance. The rotisserie, shaking platform and drip tray can be easily
removed for cleaning.
Supply
Power rating: . . . . . . . . . . . . . . . . . . . . . . . . .250 W
Frequency: . . . . . . . . . . . . . . . . . . . . . . . . .50/60 z
Fuse rating: . . . . . . . . . . . . .110/120 V: 6.3A type T
. . . . . . . . . . . . . . . . . . .220/240 V: 1.6A type T
Ingress Protection Rating . . . . . . . . . . . . . . . . .IP22
Rotisserie speed control PCB fuse: . . . . . . . .500 mA
Note This item should not be serviced by the customer.
Working Environment: . . . . . . . . . . . .15°C to 25°C
Caution Care should be taken when lifting the Oven. It is advisable that
help is sought when removing an Oven from its packaging and when
stacking Ovens for use. s
Caution During operation, caution should be taken with moving parts
that are accessible when the Oven door is open. s
Thermo offers full service and technical support for all its products.
Alternatively, contact your local authorised distributor.
Safety Precautions
Oven Temperature
Monitoring (cont’d)

Shake ‘n Stack 2-1Thermo Scientific
Section 2 Unpacking and Installation
Shake ‘n Stack Oven
4 Leveling Feet
Shaking Tray (Models 6240, 6241)
Shaking Tray Drive Arm Assembly (Models 6240, 6241)
Shaking Tray Bracket (Models 6240, 6241)
Power Cable
Drip Tray
Note If any item is missing or damaged, contact Thermo or an authorised
Thermo distributor.
Warning If it is intended to use the Oven with radioactive isotopes, the
oven must be located in a designated Radiation Area. Radiation safety
procedures must be followed at all times. s
• In the event of spillage, refer to Section 6 for guidance on cleaning and
decontamination.
• All ybridisation Oven standard rotisseries are designed to accept the
long, medium or short ybridisation Bottles.
Once unpacked, attach leveling feet to base, and position on a flat surface.
Level the oven by adjusting the height of the leveling feet. Once leveled,
the oven is ready for use. If adding to a stack of Shake ‘n Stack Ovens
(maximum of 3), attach feet and adjust height, then position feet in the 4
indents on the top of the supporting Shake ‘n Stack Oven. Some
additional adjustment of the feet may be necessary until the ovens sit in a
stable position.
Note Shake ‘n Stack Ovens have been designed to stack 3 units high. No
attempt should be made to stack higher than this.
Connect each individual oven to the mains supply using only the
removable mains lead supplied. The mains lead should be fitted with a
10 A fuse for both 110/120 V and 220/240 V regions. The oven is
grounded via the mains lead and should only be connected to an grounded
supply. (See “Technical Specifications”).

2-2 Shake ‘n Stack Thermo Scientific
Section 2
npacking and Installation
The two switches found on the control panel operate the power and the
rotisserie drive. The power switch is marked with a “1” and a “0”. By
switching to the “1” position, the mains power is connected to the oven
and the green section of the switch is exposed to indicate that the oven is
“ON”. The switch to the right operates the rotisserie/shaker. The
rotisserie/ shaker switch cannot be operated without first turning on the
power switch. To the left of the rotisserie/ shaker switch is a dial to adjust
the rotisserie speed from 5-15 rpm and the shaker speed from 4-10 rpm.
Instructions to set up the shaking platform are in Section 4.
The temperature controller has 4 buttons along the lower edge of the
controller.
When the power is switched on, the display shows three dashes (---) as the
controller goes through its internal self-tests, which are completed within 5
seconds. The display then shows the compensated temperature of the oven.
To set the controller temperature, press and release the L1 button. The L1
LED will blink on-and-off, and the display will show 1SP for 1 second,
then the present setpoint value. Press the up or down arrow button to
change the desired setpoint within the oven operating range of 35°C to
85 °C. Press the Enter button ( ) to store the new setpoint value, or press
the X button to return to normal mode without saving the new setpoint
value.
The warm up time for the oven varies according to the load and the
ambient temperature. When first switching on the oven (no bottles
installed), allow approximately one hour for the temperature to stabilise.
Solutions should be pre-warmed to the required temperature to minimise
sample stabilisation time.
Set the equired
Temperature
Figure 2-1. Temp Controller
Operation
å

Shake ‘n Stack 2-3Thermo Scientific
Section 2
npacking and Installation
For ease of use and reproducibility when hybridising in bottles, the
temperature display is calibrated to show internal bottle temperature as
opposed to oven air temperature. When the shaking platform is used,
allowance must be made for the difference between the temperature shown
on the display and the temperature achieved within a liquid-filled vessel on
the shaker.
This difference can be calculated by filling the protocol specific vessel with
enough distilled water to cover an immersible thermometer, and setting
the oven to the desired product temperature. After enough time for the
temperature to have stabilized, record the thermometer reading and
subtract this value from the setpoint value. The remainder must be added
to the setpoint value for execution of the actual protocol. For instance, a
setpoint value of 55°C minus a thermometer reading of 58°C yields a
remainder of -3°C which means that the adjusted setpoint value for this
protocol will be 52°C. Repeat this process to verify that the thermometer
reading is within +/-1°C of the setpoint value. This difference should be
determined for each oven and/or shaker protocol.
The lowest target temperature which the oven can achieve is 8°C above
ambient temperature. Avoid placing the unit in direct sunlight, or in any
area where it is likely to be exposed to hazardous or volatile chemicals.
When positioning the ybridisation Oven, ensure a gap of at least 2 in.
(5 cm) is maintained between the sides of the oven and any other
equipment or object.
The oven may be located in a fume cabinet or similar chamber, provided
the cabinet is not used to contain hazardous or volatile chemicals with low
flash points.
Full details of Technical Specifications are given in Section 8.
Note If the Shake ‘n Stack Oven is not used as specified in this manual,
the protection provided by the equipment may be impaired.
Set the equired
Temp (continued)

2-4 Shake ‘n Stack Thermo Scientific
Section 2
npacking and Installation

Section 3 Set Up the Shaking Platform
1. The shaker bracket is installed at the
factory. If replacement should ever
become necessary, instructions follow.
Continue to Step 2.
• Loosen existing screws at the bottom
of the oven back sufficiently to allow
slots in bracket to slide down onto
screw shafts. Retighten screws to
secure back plate. See Figure 2.
2. Fit the rear right hand peg of tray into
vertical slot at right of bracket,
allowing tray to temporarily rest on
oven base. Refer to Figure 3.
Shake ‘n Stack 3-1Thermo Scientific
Figure 3. Insert Tray Peg into Slot
Figure 2. Replace Shaker Bracket
Drive shaft
screw
Slot
Drive Arm
Assembly Sleeve
Fan
Oven back
Bracket
Shaker peg
Oven base
Shaker tray
Rotisserie drive shaft
Location
peg
Thumb
screw
Screw
Drive arm
Shaker peg
Figure 1. Shaking Tray Components

3-2 Shake ‘n Stack Thermo Scientific
Section 3
Set p the Shaking Platform
3. Slide drive arm assembly sleeve onto
rotisserie drive shaft, aligning
location peg within drive arm sleeve
into either slot on drive shaft,
ensuring the sleeve is pushed fully
onto shaft. Tighten using thumb
screw.
4. Locate front left peg of tray into
the hole at the end of the drive
arm.
5. Slide the rear left peg into the
horizontal slot on the left-hand side
of bracket at the back of oven.
Assembly is now complete and
ready to operate.
Figure 4. Slide Sleeve onto Shaft
Figure 5. Tray Peg into Arm Hole
Figure 6. Rear Peg into Slot

Shake ‘n Stack 4-1Thermo Scientific
Section 4 Methodology of
Hybridisation
The following procedures indicate how to use the Shake n’ Stack Oven for
hybridisation.
1. Place the bottles to be used into the oven during the warm-up period
(approximately 1 hour).
2. Select a piece of support mesh appropriate for the size of the
membrane. The recommended hybridisation mesh is supplied by
Thermo (see parts list in Section 8).
3. Pre-wet the mesh and hybridisation membrane in a suitable tray
containing 2 x SSPE (SSC). See Figure 4-1.
4. Ensuring that the hybridisation membrane exactly overlays the mesh,
roll both up into a tight roll.
If more than one membrane is to be hybridised in a bottle, simply
layer meshes and membranes as required before rolling. It is important
that each membrane is separated from another by a layer of mesh. Up
to five 0.8 x 0.8 in. (20 x 20 mm) membranes can be hybridised in a
single hybridisation bottle.
5. Place 0.34-0.51 fl. oz. (10-15 ml) SSPE (SSC) into a hybridisation
bottle and then insert the roll in such a way that the leading edge
(inside the roll) and the trailing edge are positioned relative to each
other as shown in Figure 4-3.
6. Place the bottle on a flat surface, then slowly unwind the membrane
and mesh around the inside of the bottle by gently rocking and rolling
the bottle along the surface. No air bubbles should be visible between
the membrane and the bottle. If bubbles are present, the membrane
should be removed and re-rolled.
The procedure should then be repeated more gently. Rock the bottle
backwards and forwards to attach the first part of the membrane to the
bottle. Then roll the bottle to unwind the mesh and membrane.
7. Continue until the membrane and mesh are fully unwound. The mesh
ensures the probe will have access to all parts of the hybridisation
membrane including those parts of the membrane which overlap.
Place Membranes in
a Bottle

4-2 Shake ‘n Stack Thermo Scientific
Section 4
Methodology of Hybridisation
Insert Membranes in a
Bottle
1. Assemble these items:
• Shallow tray containing 2 x SSPE (SSC).
• Mesh.
• Membrane.
Roll up mesh and membrane (Figure 4-1).
2. Place 0.3-0.5 fl. oz. (10-15 ml) of SSPE (SSC) into a bottle and then
insert the roll centrally (Figure 4-2).
3. Secure cap and, holding bottle horizontally, roll to catch the trailing
edge of the mesh; continue rolling in the same direction until coil of
mesh and membrane is well positioned.
4. Pour out SSPE (SSC) and replace with pre-warmed pre-hybridisation
buffer.
5. Place the bottle in the oven so that it rotates in the oven in the same
direction, as it was unrolled in Step 3.
Figure 4-4. Bottle in Oven Rotation
Figure 4-3. Rolling Direction of Bottle
Figure 4-2. Insert Roll
Figure 4-1. Roll Mesh and Membrane

Shake ‘n Stack 4-3Thermo Scientific
Section 4
Methodology of Hybridisation
1. Once the membrane is in place in the bottle the SSPE (SSC) can be
discarded and replaced with pre-hybridisation fluid.
This is easily done by removing the cap, pouring off the SSPE (SSC),
then pouring in the pre-hybridisation fluid. All the solutions used
during hybridisation should be pre-warmed before use. 0.34-0.51 fl.
oz. (10-20 ml) solution is recommended for medium bottles and
0.17-0.34 fl. oz. (5-10 ml) for short bottles. igher volumes will be
required if there is more than one membrane in the bottle, e.g.
0.51-0.85 fl. oz. (15-25 ml).
2. Replace the cap on the bottle and insert into the rotisserie.
1. Denature the purified probe by boiling for five minutes, then store on
ice. If the volume to be used for hybridisation is substantially different
than that used previously for hybridisation in bags or boxes, ensure
that the quantity of the probe is adjusted accordingly to maintain the
correct probe concentration. If this is not done, high background may
result.
2. Remove the bottle from the ybridisation Oven and unscrew the cap.
3. If the same buffer is to be used for hybridisation as for pre-
hybridisation, simply pipette the probe into the pre-hybridisation fluid
in the bottle. Take care to avoid pipetting the probe directly on to the
membrane as this will result in hot spots. Alternatively, dilute the
probe in pre-warmed hybridisation solution outside the bottle. Pour
off the pre-hybridisation solution and replace with the probe solution.
4. Replace the cap and gently agitate the bottle to ensure an even
distribution of the probe in the hybridisation solution.
5. Place the bottle back in the ybridisation Oven, switch on the rotisserie
and leave it to hybridise for the required time period.
Hybridisation
Pre-Hybridisation

Washing in a Sandwich
Box
Washing in Hybridisation
Bottles
Two methods for washing follow.
1. Remove the bottle from the Oven.
2. Pour off the hybridisation fluid and then half fill the bottle with the
first pre-warmed wash solution (approx. 3.38 fl. oz. [100 ml]) and
replace the cap.
Note Room temperature washing or washing with solutions which have
not been pre-warmed may result in background problems (see Appendix I
of the ybridisation Guide at the end of this manual).
3. Replace the bottle in the Oven and leave for the required time period.
If transferring from sandwich box or plastic bag methodology, the
times and temperatures are usually those currently being used,
approximately 20 minutes per wash step.
4. Repeat Steps 1-3 for each additional wash. The wash solutions,
temperatures, etc., should be those recommended by the membrane
manufacturer, or refer to the ybridisation Guide.
Note All wash solutions should be pre-warmed for best results.
Some scientists prefer to remove the membranes from the bottles and wash
them all in one container. Washing may be performed very simply in the
Shake ‘n Stack on the shaking platform and is particularly useful when
several membranes are being processed simultaneously.
1. Remove the hybridisation bottle from the oven. Pour off the
hybridisation fluid.
2. Using a pair of forceps, carefully remove the membranes and place
them in a suitable container with a lid (e.g. sandwich box or similar).
3. Using the forceps, gently unroll the membranes.
4. Add a sufficient volume of the pre-warmed first wash solution to
totally immerse the membranes. Replace the lid and shake on the
shaking platform in the Shake ‘n Stack Oven at the required
temperature for the specified time period.
Note Initial room temperature washes are not recommended and may
result in subsequent background problems.
4-4 Shake ‘n Stack Thermo Scientific
Section 4
Methodology of Hybridisation
Washing

5. Remove the first wash solution and replace it with an equal volume of
the prewarmed second wash solution. Replace the lid and shake at the
required temperature for the required time period.
6. Step 5 should be repeated for any subsequent wash. The washing
protocol using the shaking platform will be identical to that used in
the bottles.
Note Refer to Appendix I for additional information on Washing.
In addition to bulk processing of blots during stringency washing steps,
the shaking platform is ideal for performing several other stages of the blot
generation and processing procedures. Three examples are given as follows:
1. Pre-treatment of Gels prior to Nucleic Acid Transfer
Gentle agitation of the gel during pre-treatment steps is essential to
prevent damage to the gel. Pre-treatment steps are usually performed at
ambient temperature.
(i) Depurination - 0.25 M CI 10 minutes
(ii) Denaturation - 1.5 M NaCI, 0.5 M NaO , 30 minutes
(iii)Neutralisation - 1.5 M NaCI, 0.5 M Tris CI p 7.2, 30 minutes
2. Pre-washing of Filters prior to Colony Blot ybridisation
Pre-washing is often advantageous to remove colony debris and thus
reduce background noise when screening bacterial colonies by
hybridisation. After lysing the bacterial colonies and fixing the DNA
on the membrane, pre-wash in a solution of 2 x SSC/0.1% SDS at
65°C. Use several changes of buffer.
After several washes, residual colony debris may be removed by gently
rubbing filter with a gloved finger.
3. Probe Stripping from Filters after ybridisation
An example of probe stripping procedure is as follows: -
a) Wash the filter at 65°C in 5mM Tris CI p 8.0, 0.2mM EDTA,
0.1 x Denhardt’s reagent for 1-2 hours.
b) Check for residual activity by autoradiography, then re-probe.
Shake ‘n Stack 4-5Thermo Scientific
Section 4
Methodology of Hybridisation
Washing in a Sandwich
Box (continued)