Template:Kyoto/Notebook/Aug 11

From 2013.igem.org

(Difference between revisions)
(→Miniprep)
(→Colony PCR)
 
(11 intermediate revisions not shown)
Line 17: Line 17:
===Colony PCR===
===Colony PCR===
-
<!-- ここから -->
 
<div class="experiment">
<div class="experiment">
<span class="author">Kojima and Yasuda</span>
<span class="author">Kojima and Yasuda</span>
{| class="wikitable"
{| class="wikitable"
-
!Sample||base pair
+
!Sample
|-
|-
-
|BBa-B0034-(2)||-
+
|BBa_B0034 -(2)
|-
|-
-
|BBa-B0034-(1)||-
+
|BBa_B0034 -(1)
|-
|-
-
|BBa-R0011-(2)||-
+
|BBa_R0011 -(2)
|-
|-
-
|BBa-R0011-(1)||-
+
|BBa_R0011 -(1)
|-
|-
-
|negative control||-
+
|NC
|}
|}
{| class="wikitable"
{| class="wikitable"
!PreDenature||Denature||Annealing||Extension||cycle
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
|-
|-
-
|5min||30s||30s||20s||30
+
|5 min||30 s||30 s||20 s||30 cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample
 +
|-
 +
|BBa_D0412 -(1)
 +
|-
 +
|BBa_D0412 -(2)
 +
|-
 +
|NC
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
 +
|-
 +
|5 min||30 s||30 s||1 min 30 s||30 cycles
|}
|}
</div>
</div>
 +
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">Kojima anda Yasuda</span>
{| class="wikitable"
{| class="wikitable"
-
!Sample||base pair
+
!8/9 BBa_J44000 2||EcoRI||PstI||10x buffer H||MilliQ||total
|-
|-
-
|BBa-D0412-(1)||
+
|2||0.2||0.2||1||6.6||10
|-
|-
-
|BBa-D0412-(2)||
+
|2||0.2||--||1||6.8||10
|-
|-
-
|negative control||
+
|2||--||0.2||1||6.8||10
 +
|-
 +
|2||--||--||1||7.0||10
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!PreDenature||Denature||Annealing||Extension||cycle
+
!8/9 BBa_J23106||EcoRI||PstI||10x buffer H||MilliQ||total
|-
|-
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+
|1.3||0.2||0.2||1||7.3||10
|-
|-
-
|5min||30s||30s||1min30s||30
+
|1.3||0.2||--||1||7.5||10
 +
|-
 +
|1.3||--||0.2||1||7.5||10
 +
|-
 +
|1.3||--||--||1||7.7||10
|}
|}
 +
{| class="wikitable"
 +
!8/9 BBa_J23100 2||EcoRI||PstI||10x buffer H||MilliQ||total
 +
|-
 +
|1.3||0.2||0.2||1||7.3||10
 +
|-
 +
|1.3||0.2||--||1||7.5||10
 +
|-
 +
|1.3||--||0.2||1||7.5||10
 +
|-
 +
|1.3||--||--||1||7.7||10
 +
|}
 +
*incubated at 37 &deg;C for 1 hour.
 +
</div>
 +
 +
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||--||--
 +
|-
 +
|2||8/9 BBa_J44000||EcoRI||PstI
 +
|-
 +
|3||8/9 BBa_J44000||EcoRI||--
 +
|-
 +
|4||8/9 BBa_J44000||--||PstI
 +
|-
 +
|5||8/9 BBa_J44000||--||--
 +
|-
 +
|6||8/9 BBa_J23106||EcoRI||PstI
 +
|-
 +
|7||8/9 BBa_J23106||EcoRI||--
 +
|-
 +
|8||8/9 BBa_J23106||--||PstI
 +
|-
 +
|9||8/9 BBa_J23106||--||--
 +
|-
 +
|10||8/9 BBa_J23100||EcoRI||PstI
 +
|-
 +
|11||8/9 BBa_J23100||EcoRI||--
 +
|-
 +
|12||8/9 BBa_J23100||--||PstI
 +
|-
 +
|13||8/9 BBa_J23100||--||--
 +
|}
 +
[[File:Igku Aug11electrophoresis1.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||100bp ladder
 +
|-
 +
|2||BBa_B0034 (1)
 +
|-
 +
|3||BBa_B0034 (2)
 +
|-
 +
|4||BBa_R0011 (1)
 +
|-
 +
|5||BBa_R0011 (2)
 +
|-
 +
|6||NC
 +
|-
 +
|7||100bp ladder
 +
|}
 +
[[File:11 electrophoresis2.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
|1||1kb ladder
 +
|-
 +
|2||BBa_P0412 (1)
 +
|-
 +
|3||BBa_P0412 (2)
 +
|-
 +
|4||NC
 +
|}
 +
[[File:Igku Aug11electrophoresis3.jpg]]<br>
 +
</div>

Latest revision as of 07:39, 27 September 2013

Contents

Aug 11

Miniprep

Nakamoto

DNAconcentration[µg/mL]
8/9 BBa_J44000 -(2)100
8/9 BBa_J23106154
8/9 BBa_J23100144
8/10 T7-His-FT92

Colony PCR

Kojima and Yasuda

Sample
BBa_B0034 -(2)
BBa_B0034 -(1)
BBa_R0011 -(2)
BBa_R0011 -(1)
NC
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C68 °C--
5 min30 s30 s20 s30 cycles
Sample
BBa_D0412 -(1)
BBa_D0412 -(2)
NC
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C68 °C--
5 min30 s30 s1 min 30 s30 cycles

Restriction Enzyme Digestion

Kojima anda Yasuda

8/9 BBa_J44000 2EcoRIPstI10x buffer HMilliQtotal
20.20.216.610
20.2--16.810
2--0.216.810
2----17.010
8/9 BBa_J23106EcoRIPstI10x buffer HMilliQtotal
1.30.20.217.310
1.30.2--17.510
1.3--0.217.510
1.3----17.710
8/9 BBa_J23100 2EcoRIPstI10x buffer HMilliQtotal
1.30.20.217.310
1.30.2--17.510
1.3--0.217.510
1.3----17.710
  • incubated at 37 °C for 1 hour.

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
28/9 BBa_J44000EcoRIPstI
38/9 BBa_J44000EcoRI--
48/9 BBa_J44000--PstI
58/9 BBa_J44000----
68/9 BBa_J23106EcoRIPstI
78/9 BBa_J23106EcoRI--
88/9 BBa_J23106--PstI
98/9 BBa_J23106----
108/9 BBa_J23100EcoRIPstI
118/9 BBa_J23100EcoRI--
128/9 BBa_J23100--PstI
138/9 BBa_J23100----

Igku Aug11electrophoresis1.jpg

LaneSample
1100bp ladder
2BBa_B0034 (1)
3BBa_B0034 (2)
4BBa_R0011 (1)
5BBa_R0011 (2)
6NC
7100bp ladder

11 electrophoresis2.jpg

LaneSample
11kb ladder
2BBa_P0412 (1)
3BBa_P0412 (2)
4NC

Igku Aug11electrophoresis3.jpg

Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Aug_11"