Team:Kyoto/Notebook1

From 2012.igem.org

(Difference between revisions)
(August 2)
(August 2)
 
Line 14: Line 14:
|-
|-
|50||2
|50||2
-
|}  37℃,1h incubate
+
|}  37°C,1h incubate
{|class="wikitable"
{|class="wikitable"
Line 33: Line 33:
After 42°C 60sec heat shock, cells were stored on ice for 2min.<br>
After 42°C 60sec heat shock, cells were stored on ice for 2min.<br>
Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate.
Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate.
 +
<br>
 +
<br>
 +
==August 13==
 +
Liquid culture of FT
 +
<br>
 +
<br>
 +
==August 14==

Latest revision as of 13:24, 21 August 2012

August 2

Mutation of FT

Inverse PCR
10xBufer2mM dNTPsprimer fwdprimer revtemplatepolymeraseMilliQTotal
551.51.50.5135.550

94°C 2min, (98°C 10sec, 68°C 4min)x2cycles, 4°C Hold

Dpn1 Digestion
PCR productDpn1
502
37°C,1h incubate
Self-ligation
productMilliQLigaseT4 KinaseTotal
275115

16°C, 1h incubate

Transformation
competent cellDNA
202

Cells were stored on ice for 30min.
After 42°C 60sec heat shock, cells were stored on ice for 2min.
Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate.

August 13

Liquid culture of FT

August 14