Team:Colombia/Notebook/Protocols

From 2012.igem.org

(Difference between revisions)
(→PCR - Pfu DNA polimerase)
(→PCR - Pfu DNA polimerase)
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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.
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'''==
+
==='''PCR - Pfu DNA polimerase'''===
 +
 
 +
One reaction
{|
{|
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! Amount (μL)
! Amount (μL)
|-
|-
-
|H ~2~ O
+
|H2O
-
|10
+
|16.95
|-
|-
|Buffer
|Buffer
-
|4
+
|2
|-
|-
-
|MgSO_4
+
|MgSO4
-
|1
+
|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'''==
=='''PCR - Invitrogen (R) Taq polimerase'''==

Revision as of 21:51, 13 July 2012

Contents

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

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