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