Instruction manual

Table of Contents
Page
Section 1 Equipment Overview ..................................................................................1
1.1 Safety .........................................................................................................................1
1.2 Specifications .............................................................................................................2
1.3 Description of Major Components ...........................................................................3
Section 2 Introduction to Technology ........................................................................5
2.1 Overview of Denaturing Gradient Gel Electrophoresis ...........................................5
Section 3 DGE Background Information .................................................................7
3.1 Introduction ...............................................................................................................7
3.2 Perpendicular DGGE ................................................................................................8
3.3 Parallel DGGE ...........................................................................................................9
3.4 CDGE ......................................................................................................................10
Section 4 Sample and Reagent Preparation ...........................................................12
4.1 Sample Preparation .................................................................................................12
4.2 Reagent Preparation ................................................................................................14
4.3 Gel Volumes ............................................................................................................12
Section 5 Buffer Temperature ..................................................................................15
5.1 Temperature Controller ...........................................................................................15
5.2 Pre-heating the Running Buffer ..............................................................................15
Section 6 Gel Casting .................................................................................................16
6.1 Assembling the Glass Plate Sandwiches ................................................................16
6.2 Model 475 Gradient Delivery System.....................................................................21
6.3 Casting Perpendicular DGGE Gels .........................................................................21
6.4 Casting Parallel DGGE Gels ...................................................................................23
6.5 Casting CDGE Gels ................................................................................................24
Section 7 Electrophoresis ..........................................................................................26
7.1 Assembling the Upper Buffer Chamber .................................................................26
7.2 Loading Samples .....................................................................................................27
7.3 Running the Gel ......................................................................................................28
7.4 Removing the Gel ....................................................................................................28
7.5 Staining and Photographing Gel .............................................................................29
Section 8 Troubleshooting Guide .............................................................................30
8.1 Equipment ...............................................................................................................30
8.2 Applications .............................................................................................................32
Section 9 Maintenance ..............................................................................................33
Section 10 References ..................................................................................................34
10.1 Applications in Mutation Detection Electrophoresis .............................................34
10.2 Mutation Detection Electrophoresis References ....................................................39
Section 11 Instruments and Reagents for
Mutation Detection Electrophoresis .......................................................48
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