Wide Dual Mini-Verticals Mini-Vertical Slab Gel /Blotting System DCX-800C, cooled DCX-800 INSTRUCTION MANUAL
T A B L E O F C O N T E N T S Important User Information. . . . . . . . . . . . . . . . . . . . . . . . 3-4 Section 1: General Information 1.1 Introduction and Features . . . . . . . . . . . . . . . . . . . . . . . 5 1.2 Specifications and Constructions. . . . . . . . . . . . . . . . . . . . . 5 1.3. Comb Specifications for Hand Cast Gels . . . . . . . . . . . . . . . .. . 6 6 1.4 Safety. . . . . . . .
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 and Features C.B.S. Scientific offers the Wide Dual Mini-Verticals are for performing SDS-Page, acrylamide-nucleic acid separations and electro-blotting. The dual unit provides the capability of running or blotting two gels simultaneously. Units include 2 blotting cassettes, 2 foam pads, and a white plastic reservoir conversion plate to allow for single runs. The DCX-800C has a cooling base for temperature controlled runs.
1.3 Comb Specifications for Hand Cast Gels COMB OPTIONS material: PTFE, tooth depth: 9.5mm, overall length: 11.8cm Cat. # MG15-0712M* MG15-0720 MG15-0726 MG15-1012M* MG15-1020 MG15-1026 MG15-1512M* MG15-1520 MG15-1526 # of Teeth Thickness of Teeth (mm) Width of Teeth (mm) Recommended Max. Sample Vol./Well (µl)* 12+2 20 26 12+2 20 26 12+2 20 26 0.75 0.75 0.75 1.0 1.0 1.0 1.5 1.5 1.5 7.38 + 3.07 4.27 2.80 7.38 + 3.07 4.27 2.80 7.38 + 3.07 4.27 2.
SECTION 2 Description of Parts 2.1 Unpacking and Components Please verify that your unit comes complete with the following components: Wide Dual Mini-Vertical Electrophoresis System: • Lower Reservoir (DCX-800C includes cooling base with 2 tubing adapters) • Safety cover • Power leads (must be attached before using. See section 3.1.
SECTION 3 Instructions for Slab Gel Electrophoresis Using Pre-Cast Gels 3.1 Preparing the Electrophoresis Unit 1. Power leads are shipped separately and must be attached before using your Wide Dual Mini-Vertical. Thread the power lead ends into the lid receptacles and rotate in clockwise direction to hand tighten. 2. Place unit in authorized work area. Remove safety cover from the assembled unit by simultaneously pressing down on white push pins while lifting up on blue safety cover as shown in figure 1.
5. Slide pre-cast gel cassette or plate set(s) into the core assembly with the notched plate facing in towards the upper buffer reservoir as shown in figure 4. If using a pre-cast gel stored at 4°C, allow to warm to room temperature. If pouring your own gels please see Alternate Protocol (Section 4) on pages 1214 describing gel casting using Gel Wrap™. 4 6. If running one gel, slide white plastic adaptor plate into the side without the gel.
3.2 Running the Gel 1. Place stirring bar into bottom of reservoir in stirring corral (as shown in figure 7). Place core assembly into lower reservoir. The anode (red) and cathode (black) electrodes are color-coded on both the core/cassette assembly and lower reservoir. See figure 7. Ensure the red dot on the cassette assembly is on the same side as the red receptacle on the lower reservoir. Fill core upper reservoir with freshly prepared buffer (~ 190mls).
3.3 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 as shown in figure 9. Do not remove safety cover by pulling up on leads! 2. Pull up on gel door latches, and open gel door. Remove gel sandwich from Cassette Assembly. Stain and fix according to your preferred method. 11 9 www.cbsscientific.
4.1 Alternate Protocol: Using Gel Wrap™ Gasket Casting Method (Not using Pre-cast gels) 1. Place all components in an authorized work area. You will need: glass plate set, Gel Wrap™ Gasket, spacer set, comb, 3 GPC-0002 clamps, and polyacrylamide solution. Prepare and clean glass plates by hand washing both plates with a high quality lab detergent followed by a complete rinsing with dH2O. Air-dry or use a lint-free tissue.
6. Lift the assembly and stand it on the base of the clamp. 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. 7. Apply PAGE 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.
SECTION 4 Alternate Protocol for Slab Gel Electrophoresis. (Not using Pre-cast gels) 4.2 Preparing the Electrophoresis Unit when Using Gel Wrap™ Gasket Casting Method 1. Place unit in authorized work area. Remove safety cover from the assembled unit by simultaneously pressing down on white push pins while lifting up on blue safety cover as shown in figure 1. Do not remove safety cover by pulling up on leads! 1 2.
SECTION 5 Instructions for Western Blotting 5.1 Preparing the Unit for Blotting 1. Remove safety cover from the assembled unit by simultaneously pressing down on white push pins while lifting up on blue safety cover. Do not remove safety cover by pulling up on leads! Remove white core from lower reservoir by grasping core with one hand and lifting directly up. Open doors on the core assembly by pulling up on the white latches, as shown in figure 1. 1 2.
black sides 4. Insert blotting casettes into core making sure that red side faces outward. See diagram 5. 5. Close doors and re-latch by pressing down on the white latches so that assembly looks like that shown in figure 6. If running one blot, slide white reservoir conversion plate into the side without the blotting cassette. 5.2 Electro- Blotting Procedure red side 1. Place stirring bar in bottom corral of lower reservoir.
SECTION 6 Running Conditions 6.1 Recommended Power for Slab Gels: 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 reference section 6.3 for in depth discussions on practical and theoretical approaches to protein gel electrophoresis. 6.1.1 ClearPAGE Gels Run Voltage Starting Current Ending Current Approx. Run TIme 180VDC 90mA/gel 40mA/gel 30-75 minutes 6.1.
6.2 Electro-Blotting As a general recommendation, equilibrate gels (after running) with the diluted transfer buffer for 5 to 10 minutes before transfer. Blotting Buffer ClearPAGE™ Transfer Buffer 10X/20X cat. # FB82500 (see Section 6.4) 100ml (1:10 dilution) Methanol 200ml Ultrapure Water 720ml Typical Blotting Conditions for DCX-800(c) Power Supply Setting Blot Time Expected Current 6.3 200V constant 1.5 - 2.0 hours with stirring, cooling blocks 180mA / 1 gel 220mA / 2 gels References 1. 2. 3.
6.4 Recommended ClearPAGE™ Buffer Formulations As an alternative to the ClearPAGE buffers available for purchase, these formulations may be used to prepare buffers yourself. Use high-quality, low-conductance ingredients. Do NOT use acid or base to adjust the pH! Standard SDS Running Buffer, 20X for Reduced Samples (Cat. # FB60500) * pH should be between 8.4 and 8.5 at 25° C. Ingredient MW Molarity Qty/Liter Tricine (free acid) 179.17 0.8M 143.4 g Tris (free base) 121.14 1.2M 145.
6.4 Recommended ClearPAGE™ Buffer Formulations, con’t. LDS Sample Buffer, 4x, (Cat. # FB31010) *pH should be between 7.7 and 7.8 at 25° C. MW Molarity Qty/Liter Glycerol (40%) Ingredient - - 400 g Ficoll-400 (4%) - - 40 g Triethanol amine, pH7.6 149.2 0.8M 120.0 6 N HCL Lithium Dodecyl Sulfate (4%) EDTA Di-Sodium Brilliant Blue G250 (0.025%) Phenol Red Ultrapure water (fill to) 36.46 372.2 - 2mM - 93.0 g 40 g 7.44 g 0.25g 0.25 g 1000 ml Standard DNA/Native Running Buffer, 20X, (Cat.
SECTION 7 Maintenance of Equipment 7.1 Care and Handling The plastic components of the Wide Dual Mini-Vertical Electrophoresis Systems are fabricated from acrylic 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, gel tray and combs with de-ionized water.
SECTION 8 Ordering Information Wide Dual Mini-Vertical Ordering Information COMBS: See page 6. CAT.
ClearPAGE Precast Gel Selection Chart & Reagent Ordering information ClearPAGE SDS gels- 15cm(w) x 10cm(h) , packages of 10 - Cat. #’s acrylamide % 12% 4 - 12% 4 - 20% 12 well + 2 marker lanes GK-01212-10 GK-41212-10 GK-42012-10 20 well GK-01220-10 GK-41220-10 GK-42020-10 26 well GK-01226-10 GK-41226-10 GK-42026-10 CAT.
CONTACT INFORMATION Online Ordering www.cbsscientific.com Telephone Local and International: 858-755-4959 Toll Free: 800-243-4959 Sales E-mail Address sales@cbssci.com Technical Service E-mail Address technicalservice@cbssci.com Mailing Address P.O.
