Bio-Rad DCode™ Universal Mutation Detection System User Manual

Page 3

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Table of Contents

Section 1

General Safety Information ..........................................................1

1.1

Caution Symbols........................................................................................1

1.2

Precautions During Set-up .........................................................................1

1.3

Precautions During a Run ..........................................................................1

1.4

Precautions After a Run.............................................................................2

1.5

Safety ........................................................................................................2

Section 2

Introduction ...................................................................................2

2.1

Introduction to Mutation Detection Technology...........................................2

Section 3

Product Description ......................................................................2

3.1

Packing List ...............................................................................................2

3.2

System Components and Accessories.......................................................5

Section 4

Denaturing Gel Electrophoresis (DGGE, CDGE, TTGE)............11

4.1

Introduction to Denaturing Gradient Gel Electrophoresis (DGGE) ............11
Reagent Preparation................................................................................13
Gel Volumes ............................................................................................15
Sample Preparation .................................................................................16
Temperature Controller............................................................................17
Preheating the Running Buffer .................................................................18
Assembling the Perpendicular Gradient Gel Sandwich ............................18
Casting Perpendicular Gradient Gels .......................................................22
Assembling the Parallel Gradient Gel Sandwich ......................................24
Casting Parallel Gradient Gels .................................................................26

4.2

Introduction to Constant Denaturing Gel Electrophoresis (CDGE)............28
Reagent Preparation................................................................................29
Gel Volumes ............................................................................................31
Sample Preparation .................................................................................31
Temperature Controller............................................................................32
Preheating the Running Buffer .................................................................33
Assembling the CDGE Gel Sandwich ......................................................33
Casting CDGE Gels.................................................................................35

4.3

Introduction to Temporal Temperature Gradient Gel Electrophoresis
(TTGE) ....................................................................................................36
Calculating the Run Parameters ..............................................................37
Reagent Preparation................................................................................38
Gel Volumes ............................................................................................40
Sample Preparation .................................................................................40
Preheating the Running Buffer .................................................................40
Assembling the TTGE Gel Sandwich .......................................................40
Temperature Controller............................................................................41
Casting TTGE Gels..................................................................................43

Section 5

Heteroduplex Analysis................................................................44

5.1

Introduction to Heteroduplex Analysis ......................................................44

5.2

Reagent Preparation................................................................................45

5.3

Gel Volumes ............................................................................................47

5.4

Sample Preparation .................................................................................48

5.5

Temperature Controller............................................................................48

5.6

Adding the Running Buffer .......................................................................49

5.7

Assembling the Heteroduplex Analysis Gel Sandwich .............................50

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