MP Biomedicals INSB8100 User manual

INSTRUCTION MANUAL
Vacuum Blotter
Cat # INSB8100
Cat. # INSB8101
MP Biomedicals Europe MP Biomedicals North America
Tel : [phone] Tel : [phone]
Fax : [phone] Fax : [phone]
custserv.[email protected] [e-mail]
www.mpbio.com
Jul
y
. 2010

Vacuum Blotter
Description
The Vacuum Blotter consists of (see illustration on the back cover):
•Compact buffer collection reservoir with one seal and four catches for adjusting
the sealing frame
•Porous plastic divider
•Built-in regulated vacuum pump
•Command panel with (from left to right) manometer, vacuum control knob, spirit
level and On/Off power switch
•Three adjustable feet
•Safety fuse and interchangeable power supply connecting wire (on the rear)
•Brainsick (on the front)
•Adjustable frame
•Special sealing plastic sheet
The system is made airtight by adjusting the plastic sheet between the adjustable frame
and the seal placed on the body of the device. The apparatus is delivered with two
special plastic sheets to serve as masks and to guarantee an airtight seal.

Installation
Set the instrument on a horizontal work bench with the brainsick in front of you. Make
sure the Vacuum Blotter is horizontal using the spirit level and the adjustable feet.
Connect the apparatus to the power supply (220V, 50 Hz or 110V, 60 Hz depending on
the model delivered). The instrument is delivered assembled and ready for use. Please
proceed as follows when using the Vacuum Blotter for the first time.
1) Unlock the 4 catches fixing the adjustable frame.
2) Remove the adjustable frame and the plastic sealing sheets.
3) Cut out a window in a sealing sheet with dimensions matching those of the gel
containing the material to be transferred.
The side of the window should be about 5 mm smaller than the corresponding
dimensions of the gel. This 5 mm overlap is sufficient to guarantee an airtight seal. The
window may be cut out using a scalpel. It is important to avoid going too far when
cutting the corners of the window as nicks may create leaks at these points.
4) Successively place on the porous plate :

•A Whatman (Type 3MM) paper sheet to protect the membrane
•The transfer membrane (such as MP Biomedicals Positive, Neutral or HCD)
•The sealing mask taking care that it is correctly positioned on the seal (no folds)
•The gel containing the material to be transferred
Note: If the gel splits in the course of the experiment, put the gel pieces together and glue them together
by pouring some molten agarose on the splits. After cooling and gelification, a successful transfer then be
performed.
5) Set the adjustable frame in its place and place the catches on the sides to seal the
unit.
6) Cover the gel with the transfer solution of your choice (e.g. alkaline transfer or
20XSSC).
Important Note
It is not necessary to fill the whole surface within the frame with the transfer solution. It is
sufficient to cover the gel surface with a few milliliters of the solution (due to the surface tension,
the liquid will remain on the gel). These conditions ensure optimal transfer of the nucleic acids
contained in the gel. Such procedure allows you to save solutions and to monitor the good
permeation of the solution through the gel. Make sure the gel is not dried out by periodically
adding transfer solution (3-4 times). However, it is possible to fill the whole surface within the
frame cleaning will be easier if small amounts of transfer solution have been periodically added.
7) Start up the vacuum pump and allow the vacuum to establish. Adjust the vacuum
according to the type of transfer to be carried out. The knob controls the strength of
the vacuum. The knob is turned clockwise to increase the vacuum and
counterclockwise to decrease the vacuum. To prevent locking of the manometer’s
needle do not allow the needle to come down too low (no vacuum higher than 90
mbar).
During the transfer period, it is important that the gel is permanently covered with the
transfer solution. Add transfer solution periodically with a pipette.
8) End of transfer. No special precautions are needed. Switch the pump off using the
On/Off switch. Unlock the catches of the frame when the instrument is not in use to
avoid compressing the seal.

9) Cleaning. After use, discard the Whatmann paper sheet, remove the sealing mask
and the porous plate and rinse them in running water. They can be re-used.
Draining and maintenance
Pull the apparatus forward until its front foot is no longer resting on the bench. In this
position, the reservoir is tipped forward. Place a vessel under the brainsick and unscrew
its cap. Recover the transfer solutions and rinse the inside of the reservoir using a wash
bottle. Rinse regularly to avoid crystallization of saline solution inside the buffer
collection tank. If crystals form, they are difficult to remove later.
The reservoir must be regularly drained and rinsed. About 3 liters of the the buffer
solution can be collected in the trough before the vacuum pump could be affected.
Advice to Users
Conditions usually recommended for a transfer according to Southern :
•Agarose : 0.8 to 1%
•Gel size : 20 cm x 20 cm
•Gel thickness : 4 to 5 mm
•Size of fragment to transfer : up to 50 kb
•Vacuum : 40 to 50 mbar
•Time required for transfer : 45 to 60 minutes
If the vacuum is too great, the gel may be compressed, thus impeding the transfer. We
recommend that the vacuum should not exceed 55-60 mbar. If the vacuum is too low,
the transfer time may be unnecessarily extended. In extreme cases, only partial transfer
may be achieved. We advise that the vacuum should not drop below 40 mbar.
In Case of Problems
If no vacuum can be created, please check the following sources of problems:
•The sealing mask is incorrectly positioned.
•If the plastic sheet is creased above the seal or torn even slightly, air leaks are
likely to occur. Remove the frame and mask and check its positioning, particularly
at the corners of the reservoir.
•Bad positioning of the seal in its channel.
•Insufficient overlapping of the sealing mask and gel.
•The brainsick is not tight. Fasten its cap slightly.
If any other problems occur, please contact Technical Support:

[phone]
[email protected]
TECHNICAL INFORMATION
Dimensions: 330 x 100 x 410 mm
Weight: 4.5 kg
Porous transfer area: 280 x 280 mm
Power consumption: 25 W
Vacuometer: 0 to -100 mbar
Accuracy of the vacuometer: Class 1.5
ORDERING INFORMATION
Cat. # Description
INSB8100
Vacuum Blotter 220V - 50 Hz Delivered with 2 plastic masks
INSB8101
Vacuum Blotter 110V - 60 Hz Delivered with 2 plastic masks
INSB8302 Plastic masks 28x28cm (one set of five)
INSB8401 Porous Plate
INSB8402 Sealing Frame