INSTRUCTION MANUAL Adjustable Slab Gel Systems Single Units ASG-250 ASG-400 ASG-250-02 Dual Units DASG-250 DASG-400 DASG-250-02
TABLE OF CONTENTS Page Important User Information 3-4 Section 1 1.1 1.2 1.3 General Information Introduction Specifications Safety 5 5-6 7 Section 2 2.1 2.2 2.3 2.4 Description of parts Unpacking-U.S. version Components/Assembly-U.S. version Unpacking-European version Components/Assembly-European version 8 8 9 9 Section 3 3.1 3.2 3.3 3.4 3.5 3.
ENGLISH IMPORTANT USER INFORMATION This Instruction Manual will explain how to use this product safely and effectively. Please read and carefully follow the instruction manual in its entirety. The triangle/exclamation mark symbol alerts the user of the product to important operational, maintenance, and/or warranty requirements. The triangle/lighting bolt symbol alerts the user of the product to potentially hazardous electrical exposure.
FRANÇAIS INFORMATION IMPORTANTE À L'USAGE DES UTILISATEURS ESPAÑOL Le présent manuel d'utilisation explique la manière de se servir efficacement du produit en conditions de sécurité. Il est recommandé de soigneusement lire la totalité du manuel, avec ses consignes et ses instructions. El presente instructivo explica la manera de usar este producto en forma segura y efectiva. Sírvase leerlo en su totalidad y seguir detenidamente las indicaciones que contiene.
SECTION 1 General Information 1.1 Introduction The Adjustable Slab Gel Systems offered by C.B.S. Scientific are designed to meet a wide variety of applications. These systems are designed for separating and characterizing protein and can be adapted for short nucleic acid sequencing runs with the appropriate accessories. The Dual systems allow two gels to be run simultaneously under identical buffer conditions and also provides the flexibility of running gels from 145mm to 400mm (U.S. version, Cat.
Table 1 Specifications Single Adjustable Unit U.S. version ASG-250/400 adjusted to: 145mm 160mm 170mm 190mm 220mm 280mm 400mm Single Adjustable Unit European version ASG-250-02 adjusted to: 160mm 170mm 190mm 220mm 280mm Dual Adjustable Unit U.S.
1.3 Safety Power to the Adjustable Vertical units is to be supplied by an external DC voltage power supply that must be ground isolated so that the DC voltage output floats with respect to ground.
SECTION 2 Description of Parts 2.1 Unpacking—U.S. version only Please verify that your unit comes complete with the following components: Single Adjustable Vertical Gel Kit ASG-250/400: • • • • • • • • • • • • 2.
SECTION 2---continued Description of Parts 2.3 Unpacking—European version only Please verify that your unit comes complete with the following components: Single Adjustable Vertical Gel Kit ASG-250-02: • • • • • • • • • • • • 2.
Section 3 Instructions for Use 3.1 Adjustable Slab Gel Unit Preparation A. Unit Set-Up 1. Place Adjustable Slab Gel unit on a level work surface in an authorized work area. Attach each of the black guide rods to the flathead screws through the countersunk holes on the bottom of the base. Thread the screws into the black guide rods and hand tighten. Do not over tighten. 2. Slide the upper reservoir onto the guide rods.
3.2 Preparation/Cleaning of Glass Plates Hand wash both plates with a high quality lab detergent followed by a complete rinsing with dH2O. Air dry or use a lint-free tissue. Spray/wipe the chosen inner surfaces of the plate set with 95% ethanol and dry with lint-free tissue. 3.3 Gel Casting Using Gel Wrap Gasket Casting method 1. Start by holding the rectangular back plate with the rounded bottom corners and start applying the gasket around one side of the glass plate.
5. Lift the assembly and stand it on the base of the clamps. For leveling, push glass plate assembly down until it stops against clamp body. Clamp the sides of the assembly with additional casting clamps on either side. As each clamp is attached, be sure the gasket is aligned between the plates forming a seal. 6. Apply PAGE or agarose solution to gel plate sandwich using a syringe or pipette. If using a stacking gel, pour desired height of running gel, then overlay a small amount of dH2O or 0.
3.4 Running the Gel 1. Attach plate set(s) to unit with smaller or notched plate facing towards the upper buffer reservoir. If using a pre-cast gel stored at 4°C, allow to warm to room temperature. Use white clamps with mouth open (cat. # GPC-0001) to attach plate set to unit, clamping each side to the sandwich. 1b. Optional Cooling Plate Instructions: If using optional aluminum plate during electrophoresis place behind the unit shown in figure at right.
3.5 Removing the Gel 1. Turn the power supply off and disconnect the leads from the power supply. Remove the safety cover from the unit, by placing thumbs on white posts next to red & black connectors, then pushing down while pulling up with fingers under lid. DO NOT pull on power cords. 2. Drain upper reservoir using a pipette or syringe. Detach side clamps and remove gel sandwich. Carefully separate, making sure that the gel remains attached to one plate. Stain and fix according to your preferred method.
SECTION 4 Running Conditions 4.1 Recommended Power Precise electrophoresis conditions will vary according to the number and type of gels used, buffer conditions employed, power input, and the general goal of the experiment. Refer to the reference section for in depth discussions on practical and theoretical approaches to protein gel electrophoresis. Using standard SDS-PAGE buffer systems apply 1-10 VDC/cm of gel. For sequencing applications use 50 VDC/cm.
4.2 Recommended Buffers and Reagents-continued BUFFERS: Agarose TBE (1X solution): 0.089M Tris base 0.89M Boric acid 0.002M EDTA pH 8.3 Protein Denaturing TG-SDS(1X): 0.025M Tris base 0.192M Glycine 0.1%(w/v) SDS pH3 DNA Sequencing TTE (Tris/Taurine/EDTA)(1X): 1.78M Tris 0.57M Taurine 0.01M EDTA Na2-2H20 4.3 Page/Agarose Slab TAE (1X): 0.04M Tris-acetate 0.001M EDTA pH 8.0 Denaturing/Non-Denaturing/ Nylon Blotting TT (Tris-Taurine( (1X): 0.1M Tris base 0.1M Tricine TT-SDS (1X): 0.1M Tris base 0.
SECTION 5 Maintenance of Equipment 5.1 Care and Handling The plastic components of the Adjustable Slab Gel units are fabricated from acrylic, delrin and polycarbonate. Electrodes and connectors are made from pure platinum, stainless steel, and chrome plated brass. As with any laboratory instrument, adequate care ensures consistent and reliable performance. After each use, rinse buffer chamber, glass plate, spacers and combs with de-ionized water.
SECTION 6 Adjustable/Dual Adjustable Slab Gel Kits and Accessories Cat. # ASG-250 ASG-250-02 ASG-400 DASG-250 DASG-250-02 DASG-400 Item Adjustable Slab Gel Kit, U.S. only. Adjusts from 145mm to 280mm. Includes 1 comb, 1 Gel Wrap spacer set, 1 Gel Wrap Glass plate set, 1 each Gel Wrap and 8 clamps. Adjustable Slab Gel Kit-CE, adjusts from 160mm to 280mm. Includes 1 comb, 1 Gel Wrap spacer set, 1 Gel Wrap Glass plate set, 1 each Gel Wrap and 8 clamps. Adjustable Slab Gel Kit, U.S. only.
SPACERS for GEL WRAP (PVC) Cat. # Spacer dimensions GEL WRAP GASKET Cat. # Gasket thickness x plate size VGS-0512R 0.50mm x 145mm VGS-0516R 0.50mm x 160mm VGS-0517R 0.50mm x 170mm VGS-0519R 0.50mm x 190mm VGS-0520R 0.50mm x 220mm VGS-0525R 0.50mm x 280mm VGS-0540R 0.50mm x 387mm VGS-7512R 0.75mm x 145mm VGS-7516R 0.75mm x 160mm VGS-7517R 0.75mm x 170mm VGS-7519R 0.75mm x 190mm VGS-7520R 0.75mm x 220mm VGS-7525R 0.75mm x 280mm VGS-7540R 0.75mm x 387mm VGS-1012R 1.0mm x 145mm VGS-1016R 1.
STANDARD GLASS PLATE SETS, NOTCHED* GLASS PLATE SETS, NOTCHED & BEVELED* For 2-dimensional gels Cat. # Glass Plate dimensions (w x h) Cat. # Glass Plate dimensions (w x h) NGP-125N NGP-160N NGP-170N NGP-190N NGP-200N NGP-250N NGP-400N 16.5cm x 145mm 16.5cm x 160mm 16.5cm x 170mm 16.5cm x 190mm 16.5cm x 220mm 16.5cm x 280mm 16.5cm x 387mm NGP-125B NGP-160B NGP-170B NGP-190B NGP-200B NGP-250B NGP-400B 16.5cm x 145mm 16.5cm x 160mm 16.5cm x 170mm 16.5cm x 190mm 16.5cm x 220mm 16.5cm x 280mm 16.
ADDITIONAL VERTICAL GEL ACCESSORIES: BAR CLAMP Cat. # GPC-1650 SILICONE CORD REPAIR KIT Item Bar clamp for 16.5cm wide units Cat. # SCK-500 Item Kit for Sequencing and Slab Gel Units MULTIPLE GEL CASTING CHAMBERS Cat. # GCC-404 GCC-410 Item Multiple Vertical Gel Casting chamber, casts up to 4 gels simultaneously Multiple Vertical Gel Casting chamber, casts up to 10 gels simultaneously GEL CASTING STAND Cat.
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