INSTRUCTION MANUAL Mega-Gel High-Throughput Electrophoresis System C-DASG-400-50
TABLE OF CONTENTS Page Important User Information 3-4 Section 1 1.1 1.2 1.3 General Information Introduction 5 Specifications 5 Safety 5 Section 2 2.1 Description of parts Unpacking-Mega Gel 6 Section 3 3.1 3.2 3.3 Assembly Unit assembly for Mega Gel Glass Plate Preparation Gel Casting 7 7 8-9 Section 4 4.1 4.2 4.3 4.4 4.
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 1.2 Specifications This Dual Adjustable Mega-Gel is designed for high-throughput microsatellite mapping (1) with a capacity of up to 200 samples per run. The glass plate sets include a low fluorescence back plate, making it ideal for fluorescence-based assays. This feature allows the bands to be visualized without the time-consuming process of removing the gel from the glass plate sandwich and staining.
Section 2 Description of Parts 2.1 Unpacking- Mega Gel Please verify that your Mega Gel comes complete with the following components: • Buffer reservoirs, upper & divided lower with safety covers/power leads • 2 bar clamps • Rotating base with leveling feet and securing pin • 4 white nylon tubing adapters for buffer recycling • 4 knurled black thumb screws for securing lower reservoir to base • 16 white spring clamps (Cat.
Section 3 Assembly 3.1 Unit Assembly for Mega-Gel 1. Place the Mega-Gel on a level work surface in an authorized work area. Attach two guide rods to base. Tilt base on its side, place stainless steel screw through countersunk hole and screw the guide rods onto the base 2. Secure lower reservoirs to base by tightening the four black knurled thumb screws. 3. Once guide rods are secure and apparatus is flat on laboratory bench, slide the divided upper reservoir over the two guide rods.
3.3 Casting the Gel Gel Casting Using Gel WrapGasket Casting method 1. Hold the rectangular back plate (with the rounded bottom corners) and start applying the gasket around one side of the glass plate. Note: one side of the “U” shaped gasket is flat, and the other side has tubing that will act as a seal around the spacers. 2. When applying the gasket over the rounded corners of the back glass plate, make sure the notches on the gasket align with the rounded corners of the glass plate.
7. Apply gel 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 0.1% SDS solution to top of gel. After polymerization, rinse with buffer, add stacking gel solution and insert comb. For regular, single percentage gels, add polyacrylamide solution to correct height, and insert comb. Allow gel to polymerize, usually 20-30 minutes.
Section 4 Electrophoresis 4.1 Pre-electrophoresis 1. Fill lower buffer reservoir with buffer. 2. After removing the Gel Wrap Gasket from the gel sandwich, but with the white clamps still in place, rinse the sandwich in D.I. water and dry. Remove comb, then remove the white plastic clamps and place sandwich (notched plate inward) into the “buffer- filled “ lower resevoir at a slight angle to prevent capturing bubbles. Reclamp sandwich to unit using bar clamp as quickly as possible as described below. 3.
9. Connect the DC power leads to the power supply with the proper polarity. Make sure the black leads are connected to the black cathode (-) and the red leads are connected to the red anode (+). 10. Rinse sample wells with upper reservoir buffer. Before loading samples, pre-electrophorese the gel for 20 minutes. WARNING: 4.2 Excessive power will cause the gel to overheat and crack the glass plates. Loading Samples 1. At the end of the pre-electrophoresis period.
4.5 References and Suggested Reference Literature 1. Wang, Shi, Carlson, Cregan, Ward, Diers. A Low-Cost, High-Throughput Polyacrylamide Gel Electrophoresis System for Genotyping with Microsatellite DNA Markers. Crop Science (43:1828-1832) 2. MOLECULAR CLONING A Laboratory Manual, 2nd edition, ed. by J. Sambrook, E.F. Fritsch and T. Maniatis, Cold Spring Harbor Laboratory Press, 1989. 3. CURRENT PROTOCOLS IN MOLECULAR BIOLOGY ed. by F.M. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G.
Section 6 Equipment and Accessories MEGA-GEL HIGH THROUGPUT SYSTEM Cat.# Item . CDASG-400-50 Mega -Gel Vertical Kit, 50cm(w) x 22-42cm(l) Capabiltiy Kit Includes: 2ea 1.5mm x 100 well combs, 2sets 1.5mm x 22cm(l) spacers, 2sets 22cm(l) glass plates, 2ea Gel Wrap Gaskets, guide rods, 2ea Bar clamps, white spring clamps, instructions COMBS SPACER SETS Cat.
Mega-Gel Instructions 12/11 14
Mega-Gel Instructions 12/11
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