INSTRUCTION MANUAL Semi-Dry Blotting Systems EBU-3000 EBU-4000 EBU-6000
TABLE OF CONTENTS Page Important User Information 3-4 Section 1 1.1 1.2 1.3 General Information Introduction Specifications Safety 5 5 5 Section 2 2.1 2.2 Description of parts Unpacking EBU-3000, 4000 or 6000 Components/Assembly 6 6 Section 3 3.1 Instructions for Use Blotting Unit Preparation 7-11 Section 4 References 12 Section 5 5.1 5.2 Maintenance of Equipment Care and Handling Maintenance 13 13 Section 6 Equipment and Accessories 14 www.cbsscientific.
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 C.B.S. Scientific Semi-Dry Blotting Systems will transfer Western, Northern, or Southern blots reliably and quickly. These blotting systems have transfer capabilities for gel dimensions up to 12cm x 34cm (Cat. # EBU-3000), 20cm x 20cm (Cat. # EBU-4000) or 35cm x 45cm (Cat. # EBU-6000). The stainless steel cathode and platinum-coated titanium anode are durable and corrosion-resistant.
Section 2 Description of Parts 2.1 Unpacking the EBU-3000, EBU-4000 or EBU-6000 Please verify that your unit comes complete with the following components: • Blotting chamber with stainless steel cathode, and platinum-coated anode • Safety cover with power leads • Mylar sheets (for shielding uncovered portion of anode) 2.2 Components/Assembly www.cbsscientific.
SECTION 3 Instructions for Use --- The instructions are divided into the following three categories: A. B. C. D. 3.1 General Considerations Semi-Dry Electro-Blotting of DS DNA from Agarose Gels (Southern) Semi-Dry Blotting of Proteins (Western) Semi-Dry Blotting of RNA (Northern) Blotting Unit Preparation Place the blotting chamber on a level work surface in an authorized work area. A. General Considerations • Because transfer efficiency depends on many factors (e.g.
B. Semi-Dry Electro-Blotting of DS DNA from Agarose Gels (Southern) 1. Preparation for Southern Blotting a. Agarose gels up to 6mm thick may be run in these systems. Thicker gels require longer transfer times. Prior to transfer, gels should be stained with Ethidium Bromide. Ethidium bromide is a Mutagen. ALWAYS wear gloves. Dispose of solutions in accordance with the safety regulations of your institution. b. Optionally, depurinate the gel by soaking in 0.25M HCl for 20 minutes.
a. Transfer DNA of agarose gels (Southern) requires some special precautions. Excessive heat build-up can melt agarose if too much power is applied. To avoid this, try to use higher percentage gels made from high melt agarose. b. With this type of agarose, run at low current (100-200mA for 15-30 minutes, depending on the gel thickness. c. If you are using low melt agarose or low percentage agarose, use 50mA for 45-60 minutes. 5. Single stranded DNA from 6% Acrylamide a.
d. Apply a saturated membrane on the top of the paper. e. Carefully place the gel on the membrane, making sure no bubbles remain between the gel/membrane interface. f. Stack the remaining 3-4 sheets of blotting paper on the top of the gel. Add one sheet at a time and be sure to ‘smooth’ with glass rod to remove air bubbles. g. Lift the cathode assembly using the white handle in the center of lid; place lid directly over the stack and apply downward pressure while tightening each of the nylon thumb screws.
3. Preparation of the Apparatus a. Prepare 10 sheets of blotting paper cut to the same size of the gel and soak in 0.1TXAE. Cut a nylon membrane to the same size and wet in ddH2O for 5 minutes followed by equilibrate in 0.1XTAE for 20-30 minutes. b. Prepare one of the clear plastic Mylar sheets (provided in the kit) by cutting a hole in the sheet which is 1-2mm smaller than the gel dimension. c. Insert the Mylar mask over the bottom electrode anode.
SECTION 4 References 1. Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A.,Struhl, K. (ed) (1993). Current Protocols in Molecular Biology. Vol. 2, Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., Ch.10. 2. Bjerrum, O.J., Schafer-Nielsen, C., Buffer systems and transfer parameters for semi- dry electroblotting with a horizontal apparatus, Electrophoresis ’86, 315-327, VCH, Weinheim, Germany. 3. Dunn, S.D.
SECTION 5 Maintenance of Equipment 5.1 Care and Handling The plastic components of the Semi-Dry Blotting units are fabricated from acrylic and polycarbonate. Electrodes and connectors are made from pure platinized titanium, and stainless steel. As with any laboratory instrument, adequate care ensures consistent and reliable performance. After each use, rinse with de-ionized water. Wipe dry with a soft cloth or paper towel, or allow to air dry.
SECTION 6 Equipment and Accessories Cat. # EBU-3000 EBU-4000 EBU-6000 EPS-600 MM-4000 MM-6000 Item Semi-Dry Blotting System. Includes power leads and safety interlock, 12cm x 34cm. Semi-Dry Blotting System. Includes power leads and safety interlock, 20cm x 20cm. Semi-Dry Blotting System. Includes power leads and safety interlock, 35cm x 45cm. Power Supply, 96-240 VAC, 500-60 Hz, 5-600V, 3-500mA Mylar® Mask for EBU-4000, package of 4 each Mylar® Mask for EBU-6000, package of 6 each www.cbsscientific.
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CONTACT INFORMATION Telephone: Local or International 858-755-4959 Toll Free: 800-243-4959 Fax: 858-755-0733 Online ordering: www.cbsscientific.com E-mail address: sales@cbssci.com Mailing address: C.B.S. Scientific Company P.O. Box 856 Del Mar, CA 92014 Shipping address: 10805 Vista Sorrento Parkway Ste 100 San Diego, CA 92121 Credit Card Options: Visa/Mastercard Discover/American Express www.cbsscientific.
