Labnet ENDURO VE10 Manual de usuario

ENDURO™ VE10 Vertical Gel Electrophoresis Systems

User Manual

ENDURO VE10 System

E2010-PA

ENDURO VE10 Electroblotting System

E2010-PBA

Lit #M00608

Table of Contents

Page

Safety Precautions 1

Packing Lists 3

Care and Maintenance 5

Setting Up 6

Gel Casting 8

Gel Preparation 8

Gel Selection 9

Gel Pouring 10

Sample Preparation and Loading 11

Gel Running 12

Blotting Insert Setup 14

Blot Running 15

Appendix 20

Warranty 23

AccessoriesList 24

SAFETY PRECAUTION

WHEN USED CORRECTLY, THESE UNITS POSE NO HEALTH RISK.

HOWEVER, THESE UNITS CAN DELIVER DANGEROUS LEVELS OF ELECTRICITY AND ARE TO

BE OPERATED ONLY BY QUALIFIED PERSONNEL FOLLOWING THE GUIDELINES LAID OUT IN

THIS INSTRUCTION MANUAL.

ANYONE INTENDING TO USE THIS EQUIPMENT SHOULD READ THE COMPLETE MANUAL

THOROUGHLY.

The current to the unit, provided from external power supply, enters through the lid assembly,

providing a safety interlock to the user. When the lid is removed, the current to the unit is broken. DO

NOT attempt to use the unit without the safety lid correctly positioned. Always turn the power supply

off prior to removing the lid.

THE UNIT SHOULD NOT BE USED IF THERE IS ANY SIGN OF DAMAGE TO THE EXTERNAL

TANK OR LID.

ACRYLAMIDE IS A POWERFUL NEUROTOXIN IN SOLUTION FORM.

POLYMERIZED GELS CAN CONTAIN SOME UNPOLYMERIZED SOLUTION AND PROTECTIVE

GLOVES, SAFETY GLASSES AND CLOTHING MUST BE WORN.

THESE UNITS COMPLY WITH THE STATUTORY CE SAFETY DIRECTIVES:

73/23/EEC: LOW VOLTAGE DIRECTIVE: IEC 1010-1:1990 plus AMENDMENT 1:1992

EN 61010-1:1993/BS EN 61010-1:1993

Important Notice

Care and Maintenance:

These products and associated accessories should never come into

contact with the following cleaning agents, as these will cause

irreversible and accumulative damage:

Acetone

Chloroform

Methanol

Carbon tetrachloride

Phenol

Alkalis

Ethanol

Isopropyl alcohol

Water at temperatures above 60°C will cause damage to the acrylic

tanks, trays and other parts.

The tanks should be thoroughly rinsed with warm or distilled water but vigorous

cleaning is not necessary or advised. Air drying is recommended before use.

Cleaning Products and Accessories

Products are best cleaned using warm water and a mild detergent.

The units should only be cleaned with the following:

Warm water with a low concentration of soap or other compatible mild detergent.

Compatible detergents include dishwashing liquid, Hexane and Aliphatic

hydrocarbons.

The units should not be left in detergents for more than 30 minutes

PACKING LISTS:

ENDURO VE10 System

Units include tank, lid, internal module and electrodes and include the following

accessories:

Glas

s Plates

Combs

Casting

Base

Cooling

Pack

Cables

E2010

VE10 Vertical

Gel System

(E2110-NG-2) Glass

Plates, Notched, Pk/2

(E2110-PG-1-BS) Glass

Plates, Plain with bonded

1mm spacers, Pk/2

(E2110-DP) Dummy Plate

(E2110-12-1)

qty of 2, 1mm

thick, 12 sample

combs

E2110-PC E2110-CP E1107-EP

ENDURO VE10 Electroblotting System

Units include tank, lid, internal module, electrodes, Blotting module and include the

following accessories:

Glass Plates

Combs

Casting

Base

Cooling

Pack

Cables

ENDURO VE10

Electroblotting

System

(E2110-NG-2) Glass

Plates, Notched, Pk/2

(E2110-PG-1-BS) Glass

Plates, Plain with

bonded 1mm spacers,

Pk/2

(E2110-DP) Dummy

Plate

(E2110-12-1)

qty of 2, 1mm

thick, 12 sample

combs

E2110-PC E2110-CP E1107-EP

Electroblotting

module

(E2110-B-MC) 4 Blotting Cassettes, (E2110-B-FP) Pack of 8 Fiber pads

The packing lists should be referred to as soon as the units are received to

ensure that all components have been included. The unit should be checked

for damage when received. If damaged, contact carrier and save the box for

inspection. Please contact Labnet International if there are any problems or

missing items.

Usage Guidance and restrictions:

• Maximum altitude 2,000m.

• Temperature range between 4°C and 60°C.

• Maximum relative humidity 80% for temperatures up to 31°C decreasing linearly to

50% relative humidity at 40°C.

• Not for outdoor use.

This apparatus is rated POLLUTION DEGREE 2 in accordance with IEC 664.

POLLUTION DEGREE 2, states that: “Normally only non-conductive pollution

occurs. Occasionally, however, a temporary conductivity caused by condensation

must be expected.”

Care and Maintenance:

Cleaning ENDURO VE10 System

Units are best cleaned using warm water and a mild detergent. Water at

temperatures above 60°C can cause damage to the unit and components. The

tank should be thoroughly rinsed with warm water and distilled water to prevent build

up of salts but care should be taken not to damage the enclosed electrode.

Vigorous cleaning is not necessary or advised. Air drying is recommended before

use.

The units should never come into contact with the following cleaning agents,

these will cause irreversible and cumulative damage:-

Acetone, Phenol, Chloroform, Carbon tetrachloride, Methanol, Ethanol, Isopropyl

alcohol, Alkalis.

RNase Decontamination

This can be performed using the following protocol:-

Clean the units with a mild detergent as described above.

Wash with 3% hydrogen peroxide (H2O2) for 10 minutes.

Rinse with 0.1% DEPC- (diethyl pyrocarbonate) treated distilled water,

Caution: DEPC is a suspected carcinogen. Always wear gloves and safety

glasses.

RNaseZAP™ (Ambion) can also be used. Please consult the instructions for use

with acrylic gel tanks.

Questions and Service:

Should you have a question about the operation of the Spectrafuge Mini Centrifuge or

if service is required, contact Corning at: 800-492-1110. Do not send in a unit for

service without first calling to obtain a repair authorization number. Should the unit

require return to Corning for service, it should be properly packed to avoid damage.

Any damage resulting from improper packaging shall be the responsibility of the

user.

Setting up the ENDURO VE10 Gel Tanks:

Instructions for installing Electrical Leads.

1. Note the position of the lid on the unit. This shows the correct polarity and the

correct orientation of the cables, black is negative and red positive.

2. Remove the lid from the unit. Note if the lid is not removed, fitting the cables may

result in un-tightening of the gold plug and damage to the electrode.

3. Screw the cables into the tapped holes in the lid as fully as possible so that there

is no gap between the lid and the leading edge of the cable fitting.

4. Refit the lid.

The unit is now ready for use.

Vertical Gel Casting Using the ENDURO VE10 Gel Casting System:-

1. Clean a set of glass plates for each gel first with distilled water and then with

70% ethanol. One set of glass plates constitutes one notched glass plate and

one plain glass plate with bonded spacers. When using a triple glass plate

sandwich, two notched glass plates, one set of free spacers and a set of plain

glass plates with bonded spacer. The plain glass plate is positioned outermost,

then a notched glass plate, free spacers and second notched glass plate.

Alternatively, accessory notched glass plates with bonded spacers are available.

All glass plates, modules and casting base accessories must be completely

dry for setup. Wet components are more likely to misalign and cause leaks.

2. Assemble the glass plates so that the bottom of the glass plates and the spacers

are perfectly aligned. For triple plate sandwiches, the free spacers need to be

perfectly aligned which is best performed using a small spacer or comb to push

the spacers apart. Notched glass plates with bonded spacers do not need

manual alignment. NOTE: The spacers bonded to the glass plates are

slightly longer than the plates and marked with an arrow to indicate the top.

3. The ENDURO VE10 Gel insert contains pressure bars which impart even

pressure onto the edge of the glass plate. Ensure that the pressure bars are

adequately open for the thickness of spacer used. The bar can be opened by

sliding open clamps. When using a triple glass plate sandwich, the pressure bars

will need to be in the completely open position.

4. Position the ENDURO VE10 Gel module on a flat surface. Do not insert the

ENDURO VE10 Gel module into the casting base at this point.

5. Place the glass plate/spacer assembly into the Vertical Gel Insert between the

pressure bar and the blue gasket. Check that the bottoms of the glass plates are

touching the bench and fully tighten the sliding clamps. When only one gel is

being run, the dummy plate must be used in the second position and fully

tightened. NOTE: Be sure that the glass plates and the bottom of the

spacers are evenly aligned.

6. Open the cam handles on the casting stand and position the cams so they face

downwards. Position the ENDURO VE10 Gel insert on the gasket of the casting

stand so the key holes are facing the cams. The top of the ENDURO VE10 Gel

insert will need to be pushed down very slightly to insert the cams.

7. With the cam handles facing directly downwards, rotate the cams in the opposite

direction 1800 or until the insert has tightened down onto the gasket. Do not

overturn as this will cause the glass plates to push upwards and the

assembly will be more likely to leak.

NOTE: Always reverse the gasket after casting to avoid indentations. Do not

leave glass plates tightened into the casting stand for long periods of time as

this can cause damage the silicone mat. The unit is now ready for gel

preparation.

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