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Bio-Rad MicroPulser™ Electroporator User Manual

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Table of Contents

Page

Section 1 Safety Information.......................................................................................1

1.1 Electrical

Hazards.......................................................................................................1

1.2 Mechanical

Hazards ...................................................................................................1

1.3

Other Safety Precautions............................................................................................1

Section 2 Introduction ..................................................................................................2

2.1

Overview of electroporation theory...........................................................................2

2.2

Manipulation of instrument parameters.....................................................................4

Section 3 Factors Affecting Electroporation .............................................................5

3.1 Cell

growth .................................................................................................................6

3.2 DNA............................................................................................................................6
3.3 Electroporation Media................................................................................................7

Section 4 MicroPulser Operating Instructions..........................................................9

4.1

Setting up the MicroPulser System............................................................................9

4.2

Operation of the MicroPulser.....................................................................................9

4.3

Electroporation using the MicroPulser ...................................................................11

Section 5 High Efficiency Electrotransformation of E. coli...................................12

5.1

Preparation of electrocompetent cells......................................................................12

5.2 Electroporation .........................................................................................................12
5.3

Solutions and reagents for electroporation ..............................................................13

Section 6 Electroporation of Staphylococcus aureus...............................................13

6.1

Preparation of electrocompetent cells......................................................................13

6.2 Electroporation .........................................................................................................14
6.3

Solutions and reagents for electroporation ..............................................................14

Section 7 Electroporation of Agrobacterium tumefaciens .....................................15

7.1

Preparation of electrocompetent cells......................................................................15

7.2 Electroporation .........................................................................................................15
7.3

Solutions and reagents for electroporation ..............................................................16

Section 8 Electroporation of Saccharomyces cerevisiae..........................................16

8.1

Preparation of electrocompetent cells......................................................................16

8.2 Electroporation .........................................................................................................16
8.3

Solutions and reagents for electroporation ..............................................................17

Section 9 Electroporation of Schizosaccharomyces pombe.....................................17

9.1

Preparation of electrocompetent cells......................................................................17

9.2 Electroporation .........................................................................................................18
9.3

Solutions and reagents for electroporation ..............................................................18

Section 10 Electroporation of Dictyostelium discoideum ..........................................18

10.1

Preparation of electrocompetent cells......................................................................18

10.2 Electroporation .........................................................................................................19
10.3

Solutions and reagents for electroporation ..............................................................19

Section 11 Electroporation of Pichia pastoris ............................................................19

11.1

Preparation of electrocompetent cells......................................................................19

11.2 Electroporation .........................................................................................................20
11.3

Solutions and reagents for electroporation ..............................................................20