Bio-Rad Whole Gel Eluter and Mini Whole Gel Eluter - Manual (Page 2)

Table of Contents
Page
Section 1 General Information....................................................................................1
1.1 Introduction ................................................................................................................1
1.2 Specifications .............................................................................................................1
1.3 Safety ..........................................................................................................................2
Section 2 Description and Assembly of Components ...............................................3
2.1 Whole Gel Eluter Components ..................................................................................4
2.2 Vacuum Harvester Components ................................................................................5
Section 3 Assembly and Operation.............................................................................6
3.1 Preparative Slab Gel Electrophoresis.........................................................................6
3.2 Whole Gel and Mini Whole Gel Eluter Assembly....................................................6
3.3 Running the Whole Gel Eluter.................................................................................10
3.4 Harvesting Eluate .....................................................................................................10
Section 4 Optimizing Conditions ..............................................................................12
4.1 Buffer Selection........................................................................................................12
4.2 Running Conditions .................................................................................................13
4.3 Optimization Procedure ...........................................................................................14
Section 5 Maintenance and Sterilization..................................................................14
5.1 Cleaning the Whole Gel Eluter ................................................................................14
5.2 Sterilization of the Whole Gel Eluter.......................................................................14
5.3 Cleaning the Vacuum Harvester ..............................................................................14
Section 6 SDS-PAGE ..................................................................................................15
6.1 Reagents for SDS-PAGE Slab Gels (Laemmli buffer system) 3 .............................15
6.2 Preparing SDS-PAGE Gels of any %T ...................................................................16
6.3 Separating Gels - Calculating %T............................................................................17
6.4 Stacking Gel - 4%T (0.125 M Tris pH 6.8).............................................................17
Section 7 Native PAGE ...............................................................................................18
7.1 Native PAGE Slab Gels ...........................................................................................18
7.2 Reagents for Discontinuous Native PAGE (Ornstein-Davis) .................................18
7.3 Prepare Ornstein-Davis Acrylamide Gels ...............................................................19
7.4 Reagents for Continuous Native PAGE ..................................................................19
7.5 Prepare Continuous Gels (10 ml acrylamide monomer solution)...........................21
7.6 Sample Preparation ..................................................................................................21
Section 8 Troubleshooting Guide ..............................................................................21
Section 9 Equipment and Accessories ......................................................................22
Section 10 References ...................................................................................................22