"

From 2012.igem.org

Contents 1 Protocols 1.1 Bacterial DNA extraction protocol 1.2 Miniprep 1.3 Electroporation 1.4 PCR - Pfu DNA polimerase 1.5 PCR - Invitrogen (R) Taq polimerase

Protocols

Bacterial DNA extraction protocol

Miniprep

Electroporation

Electro-competent cells of E. coli DH5α are electroporated at 1.25 mV, immediately we add SOC medium to the bacteria and we incubated on a shaker at 37°C for one hour, then, we culture cells in a solid medium containing the antibiotic of selection.

PCR - Pfu DNA polimerase

One reaction

Reactives	Amount (μL)
H2O	16.95
Buffer	2
MgSO4	1.6
dNTP's	0.4
Primer Fw	0.4
Primer Rv	0.4
Taq	0.06
Pfu	0.19
DNA	2
Total	10

PCR - Invitrogen (R) Taq polimerase