Download Mini-Gel Electrophoresis Unit for DNA, RNA and Protein

SUBSIDIARIES
BELGIUM
EUROGENTEC s.a.
Tel. 04 372 74 00
Fax 04 372 75 00
FRANCE
EUROGENTEC France s.a.
Tel. 02 41 73 33 73
Fax 02 41 73 10 26
SWITZERLAND
EUROGENTEC s.a.
succursale de Genève
Français:
Tel. +32 4 366 61 00
Fax +32 4 264 07 88
Deutsch:
Tel. +49 221 258 94 55
Fax +49 221 258 94 54
i-MUPID
GERMANY
EUROGENTEC Deutschland GmbH
Tel. 0221 258 94 55
Fax 0221 258 94 54
THE NETHERLANDS
EUROGENTEC Nederland b.v.
Tel. 043 363 40 37
Fax 043 363 77 65
Ref. GEN-iMUPID-MANUAL-0803-V4
Mini-Gel Electrophoresis Unit
for DNA, RNA and Protein
Parc scientifique du Sart Tilman • 4102 SERAING • BELGIUM
Tel.: +32 4 372 74 00 • Fax: +32 4 372 75 00
[email protected] • www.eurogentec.com
Consumables
Agaroses
Molecular Biology Grade
Small Fragments
Nucleic acid length range
> 1000 bp
< 1000 bp
Gelling temperature*
37 ~ 39 C
29 ~ 33 C
Melting temperature
88 ~ 90 C
Gel strength*
1500 g/cm2
Introduction _________________________________________3
Components _____________________________________ 4 - 5
Setup / warranty ____________________________________6
Instructions for use ___________________________________7
Set the i-Mupid
__________________________________7
DNase or RNase activity
DNA binding
Electroendosmosis
ND
ND
ND
0.1 ~ 0.2
0.06 ~ 0.14
0.1 %
Buffers____________________________________________8
Agarose ________________________________________8
1000 g/cm2
ND
Sulfate
Gels _____________________________________________9
85 C
0.10 %
* measured in 1.5% agarose solution
Agarose "Molecular Biology Grade"
100 g
500 g
1 Kg
50 g
100 g
Agarose "Small Fragments"
EP-0010-01
EP-0010-05
EP-0010-10
EP-0020-05
EP-0020-10
Polyacrylamide _____________________________10 - 11
Storage _______________________________________11
Electrophoresis _____________________________________12
SmartLadder
The SmartLadder is a ready-to-use molecular weight marker, specially designed for easy quantification as
well as size determination.
Gel _____________________________________________12
Band size
Buffer ___________________________________________12
Sample Loading __________________________________13
Accessories ________________________________________14
Consumables ______________________________________15
ng/band
10000
100
8000
80
6000
60
5000
50
4000
40
3000
30
2500
25
2000
20
1500
15
1000
100
800
80
600
60
400
40
200
20
Band size ng/band
1000
800
SmartLadder SF
2
80
700
600
60
500
400
40
300
200
20
100
SmartLadder SF
SmartLadder
SmartLadder
100
400 lanes
1000 lanes
200 lanes
MW-1700-04
MW-1700-10
MW-1800-02
15
Accessories
i-Mupid
The Maxi-Plate allows you to cast fullsize gels, and to take advantage of
the complete capacity of the i-Mupid.
With these 10 x 11cm gels and the
two combs that are delivered with it,
you can run up to 48 samples at the
same time.
The Maxi-Plate allows agarose gels
only.
Size: 10x11 cm
Delivered with 2 double combs
Maxi-Plate for Mupid-2, -21 and i-Mupid
MU-0000-MP
The Smart-Comb set consists of two combs which fit into either the
regular gel maker or into the Maxi-Plate (see above). These combs
allow you to make slots of four different sizes.
Smart-Comb set for Mupid2, -21 and i-Mupid
MU-0000-SC
14
The Mupid is an ultra-compact electrophoresis system, which breaks
with existing trends. It has drawn widespread attention for its
convenience and ease of use, and has been used successfully by
researchers wordwide.
The well-known Mupid-21 has been redesigned and upgraded to the
i-Mupid, which now allows the researcher to perform agarose and
polyacrylamide gel electrophoresis using a substantially improved
power supply.
The i-Mupid is a novel and useful device for use in molecular biology
and diagnostic labs. User responses have been universally favorable.
DANGER! HIGH VOLTAGE!
• The i-Mupid should always be operated with caution. Careless handling may
result in electrical shock.
Never operate damaged or leaking equipment.
• Always turn off and remove the power supply before removing the cover.
• Certain reagents mentioned in this manual are of a hazardous nature (e.g.
ethidium bromide, acetic acid and boric acid). The researcher is cautioned to
exercise care with these reagents and with the equipment (e.g. ultraviolet (UV)
lamps, electrophoresis apparatus and high-voltage power supplies) mentioned
in this manual. Always follow the manufacturer’s safety recommendations.
• Never open the voltage transformer or the power supply. If pieces or cables are
broken or damaged, never replace them yourself.
• Never handle the electrical units with wet hands.
• Always keep high-voltage equipment away from heat and liquids.
3
i
Sample Loading
Gel maker set (ref. MU-000S)
Comb
Cover for
PAGE
This set is capable of producing 6 gels
(2 large and 4 small) at once. It may also be
used for gel storage.
Gel Maker
Plate (Small)
Specifications:
Gel Maker
Plate (Large)
Dimensions: 240 x 28 x 126 mm (WxHxL)
Maximum output: 2 large and 4 small gels
Gently load the the sample into the well using a micro pipette or micro syringe.
l Adding small volumes of glycerol or sucrose to the sample facilitates sample.
l The maximum loading volume of sample is about 10 l for a 4 mm thick gel.
l Gel fragments can be easily washed out of the well with a micro pipette, syringe or
Pasteur pipette.
Note:
Note : The cover plate is meant for PAGE gels
and should not be used for agarose gels.
Gel Maker Stand
Do not put your fingers in the solution during the electrophoresis run.
Power supply
Low-cost, compact and lightweight.
These features have been achieved by simplifying the circuitry.
Easy to operate.
Either 35, 50, 100 or 135 V DC can be selected as the output
voltage. These voltages are sufficient for ordinary
electrophoresis.
Place the cover as shown.
Reliable.
The simplified circuitry ensures a long period of trouble-free,
stable operation.
This power supply works on pulsating DC current (Pat. pending).
This allows resolutions comparable to that run by smooth DC
current. In general, a poor separation is due to uneven local
gradient potentials, caused by an uneven gel.
Dimensions: 44 x 52 x 1115 mm
(W x H x L)
Input:
100-240 V AC, 50-60 Hz
Output:
35, 50, 100 or 135 V DC
Fuse:
Automatic return type
4
13
Electrophoresis
Gel
Mini-gel migration tank (not available separately)
1. Holding the vertical portions of the plate, remove the plate containing the gel from the
stand.
If it is difficult to remove the gel maker plate (for example, if gel is trapped under the base
plate), gently push the vertical portions inward.
Ease of operation through the directly coupled
power supply.
Main Unit Cover
Both nucleic acids and proteins can be analyzed.
Migration
distance index
line
Gels prepared with the gel maker set can be set
together with the gel maker plate.
Lines with 5 mm spacing are drawn on the gel
maker plate, so that the approximate migration
distance can be determined at a glance.
Gel Marker plate
(large)
Water level
reference
For safety reasons, the power supply can not be
inserted unless the lid covers the migration tank.
2. Place the gel maker plate on the gel bed in the center of the migration tank.
The migration tank accommodates one large or two small plates.
Specifications:
Migration tank
Dimensions: 199 x 75 x 137 mm (W x H x L)
Gel size: one large gel measuring 107 x 60 mm (W x L) or two small gels measuring 52 x 60 mm
(W x L) can be cast. The electrical circuit is closed by the main unit cover.
Buffer
1. Pour buffer in the migration tank to a level slightly (3 to 5 mm) above the gel surface.
NOTE: If the cover is clouded, you can clear it by applying a few drops of UF-I on the underside of
the lid and wiping the solution evenly over the surface. Re-apply whenever necessary.
i-MUPID Electrophoresis system
l
l
MU-0030
The buffer level should be below the reference mark on the migration tank. lf the gel
maker plate is completely submerged, the migration pattern will be affected.
The buffer level reference mark may also be used for verifying that the migration tank is
level.
12
5
Set-up
1- Unpack and carefully examine the power supply, the electrophoresis chamber and the gel
maker.
6. After the gel has set, pour a little buffer solution around the comb and slowly draw out
the comb. Do not remove the plate for PAGE. The plate should remain attached to the
gel.
2- Report any damage to Eurogentec and save all packing material if any part is found to be
damaged during transport.
3- Do not attempt to use this apparatus if it is damaged.
4- This apparatus has been designed to be connected with an electrophoresis power supply.
l
5- Do not attempt to use the apparatus in any of the following cases:
l
• Buffer leaks from the main chamber.
• The electrophoresis chamber or the safety cover are cracked.
• Any of the electrical cables are worn or frayed.
l
The degree of gel hardening can be judged from the state of acrylamide between the
comb teeth.
The time required for polymerization varies with the volume and temperature of the gel
solution.
The polymerization time is normally 30 to 60 minutes, although longer times (2 to 3
hours at 25 C) are ideal.
Caution:
6- Place the apparatus close to its electrophoresis power supply. Make sure that this is a safe
and dry location.
7- Please refer to the instruction summary for details on how to set up the i-Mupid.
Allow the gel to solidify completely. If the gel has not yet completely set or if the comb is
withdrawn (too) rapidly, wells may be damaged and/or gel fragments may drop into the
well. The resulting bands will be distorted.
8- Warranty 1 year (replacement fuse excluded).
Note:
• It is important to keep the gel unit clean, particularly the gel maker plate, cover and
comb. We recommend that the gel maker set be cleaned with a sponge moistened with
detergent, washed in water, and then allowed to dry.
• Do not clean any part of the gel unit with organic solvents such as ethanol or acetone.
The unit will be damaged by organic solvents and any guarantees will be void.
Gel storage
If you wish to store gels you have prepared,
• Be sure to cover the gel with buffer.
• Store the gel at a low temperature
• Avoid prolonged storage.
6
11
Instructions for use
i
Polyacrylamide gel preparation
Set the i-Mupid
1. Place the gel maker stand horizontally on the bench.
2. Place the gel maker plate on the gel maker stand. Good results have been obtained by
covering the iside of the gel maker plate with a gel bonding sheet. Cut the sheet to the
specified size (106 58 mm for a large gel, 51 58 mm for a small gel). These sheets
protect the polyacrylamide gel and facilitate handling.
3. Gently pour the de-gassed acrylamide/SDS acrylamide solution between the gel maker
plates using a pipette.
l The cover included with the system will produce 3 mm-thick gels. About 24 ml of
acrylamide is needed for the large gel maker plate and about 12 ml for the small gel
maker plate.
l If air is trapped between the gel maker stand and the plate, the plate may tilt. To
dislodge the air bubble, push the gel maker plate using a pipette or press clown on the
vertical portion of the plate. Ensure that no air clings to the gel surface.
4. Place the bottom end of the plate for PAGE in the acrylamide solution in the gel maker
plate (see figure). If air bubbles are trapped under the cover, raise the other end and then
lower it again to dislodge air bubbles.
l
Acrylamide can be added through the cover slit for the comb after the gel bond sheet
and plate for PAGE are positioned on the gel maker plate. To do this, slowly pipette the
acrylamide solution, while moving the pipette from one end of the slit to the other.
It is recommended that after the cover is set over the acrylamide solution, acrylamide
be added to slightly above the cover slit (as shown below).
You only want to set the voltage
• Press power on
• Set with the “voltage” selector the desired voltage (35, 50, 100 or
135Volts). A green led shows you the selected voltage.
• To run the migration, press the “Set” selector (when the power is
running a green flickering light can be seen)
• To stop the run, press the “power off” selector
You want to use your i-Mupid with the timer
• Press power on
• Set with the “voltage” selector the desired voltage (35, 50, 100 or
135Volts). A green led shows you the selected voltage.
• To set the running time press the “+” selector. To increase the time
use the “+” selector (maximum 99min). To decrease the time use
the “-“ selector (minimum 1min).The timer always start at 30min.
• To start the migration, press the “Set” selector (when the power is
running a green flickering light can be seen)
• A timing countdown can be seen At the end of the selected time the
migration is automatically stopped and a bell is heard.
Fuse
The i-Mupid has got an automatic return type fuse. The circuit could be cut in any
abnormal condition (eg heating up condition due to a too concentrate buffer). To
reset the power supplyer, you have to remove the electric source cord for at least
30 seconds. After that period, the circuit will return to normal condition. You have
to change the buffer with a 0.5 X TBE or TAE.
5. Insert the comb through the cover slit. This is easily accomplished by slightly shifting
either the comb or the cover.
10
7
Migration buffer and its temperature
Some optimum buffer solutions are given below:
If the buffer temperature rises above 40 C during electrophoresis, turn off the power and either
dilute the running buffer and/or immerse the longer half of the Migration Tank in an ice bath.
Alternatively, the i-Mupid can be used in a cold room.
Agarose gel preparation - Preparing gels with the gel maker set
For DNA
Agarose gels:
(cf. p.15)
Polyacrylamide gels:
Tris
EDTA 2Na
Acetic acid
Add distilled water to make
4.8 g
0.74 g
1.14 ml
1 l (pH 8.1)
Tris
60.6 g
Boric acid
41.5 g
EDTA 2Na
37.2 g
Add distilled water to make 1 l (pH 8.3) for storage.
Dilute the stock solution 1:10 with distilled water before use.
For proteins
Agarose gels:
(cf. p.15)
1. Place the gel maker stand in a horizontal position.
2. Install the gel maker plate on the gel maker stand. (Note: the cover for PAGE is not
required.)
One of the ends of the gel maker plates is gray.
This is the end where the comb will be placed.
The gray color will improve the visibility of the slots.
3. Set the comb.
4. Allow the molten agarose (cf. p. 15) to cool to below 70 C before pouring, to prevent
warping of the gel maker plate.
5. Pour the gel. For 4 mm thick gels, about 25 ml of agarose (cf. p. 15) is required for the large
gel maker plate and 15 ml for the small gel maker plate. The gel thickness can be modified as
needed (see below).
Tris
6.0 g
Glycine
28.8 g
Add distilled water to to make 1 l (pH 8.3) for storage.
Dilute the stock solution 1:10 with distilled water before use.
SDS polyacrylamide gels:
Buffer 1:
Tris
15.0 g
Glycine
72.5 g
Add distilled water to make 500 ml (pH 8.3) for storage.
Dilute the stock solution 1:10 with distilled water before use.
Buffer 2:
NaH2PO4 • 12 H20
35.8 g
NaH2PO4 • H20
3.9 g
SDS
1.0 g
Add distilled water to make 500 ml (pH 7.1) for storage.
Dilute the stock solution 1:30 with distilled water before use.
8
(Effective thickness)
If air is trapped between the gel maker stand and the plate, the plate may tilt. To dislodge
the air bubble, push the gel maker plate using a pipette or press down on the vertical
portion of the plate. Ensure that no air clings to the gel surface.
6. After the gel has set, pour buffer solution onto the gel to a level just above the gel
surface and gently draw out the comb.
Caution:
Allow the gel to solidify completely. If the gel has not yet completely set or if the comb is
withdrawn (too) rapidly, wells may be damaged and/or gel fragments may drop into the
well. The resulting bands will be distorted.
9