Team:Paris Saclay/Notebook/July/16

From 2013.igem.org

(Difference between revisions)
Line 18: Line 18:
Anaïs
Anaïs
 +
* DNA : 5µL
* DNA : 5µL
* Buffer FD : 2µL
* Buffer FD : 2µL
Line 69: Line 70:
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]
 +
 +
==='''B - PCB sensor system'''===
 +
 +
===='''Objective : obtaining Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3'''====
 +
 +
===='''1 - Electrophoresis of the digestion of Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3'''====
 +
 +
Anaïs, Zhou
 +
 +
* Bba_K1155001 :
 +
 +
{|
 +
| style="width:350px;border:1px solid black;" |[[]]
 +
| style="width:350px;border:1px solid black;vertical-align:top;" |
 +
* Well 1 to 5 : 2µL Bba_K1155001 digested by EcoRI/PstI+2µl of 6X loading dye
 +
* Well 6 and 7 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
 +
* Well 8 : 6µL DNA Ladder
 +
* Gel : 1.2%
 +
|}
 +
 +
Expected size :
 +
* Bba_K1155001 digested by EcoRI/PstI : 2037bp + 333bp
 +
* Bba_K1155001 digested by SacII : 2370bp
 +
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DE;" |
 +
In the first well, we obtain fragment at the right size. We will amplify Bba_K1155001.
 +
|}
 +
 +
* Bba_K1155002 :
 +
 +
{|
 +
| style="width:350px;border:1px solid black;" |[[]]
 +
| style="width:350px;border:1px solid black;vertical-align:top;" |
 +
* Well 1 to 8 : 2µL Bba_K1155002 digested by EcoRI/PstI+2µl of 6X loading dye
 +
* Well 9 : 6µL DNA Ladder
 +
* Well 10 to 17 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
 +
* Gel : 1.5%
 +
|}
 +
 +
Expected size :
 +
* Bba_K1155002 digested by EcoRI/PstI : ...
 +
* Bba_K1155002 digested by SacII : ...
 +
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DE;" |
 +
We didn't obtain fragments at the right size. We will do the ligation again.
 +
|}
 +
 +
* BphR2 in PSB1C3 :
 +
 +
{|
 +
| style="width:350px;border:1px solid black;" |[[]]
 +
| style="width:350px;border:1px solid black;vertical-align:top;" |
 +
* Well 1 to 8 : 2µL Bba_K1155002 digested by EcoRI/PstI+2µl of 6X loading dye
 +
* Well 9 : 6µL DNA Ladder
 +
* Well 10 to 17 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
 +
* Gel : 1.5%
 +
|}
 +
 +
Expected size :
 +
* Bba_K1155002 digested by EcoRI/PstI : ...
 +
* Bba_K1155002 digested by SacII : ...
 +
 +
{|
 +
| style="border:1px solid black;padding:5px;background-color:#DE;" |
 +
We didn't obtain fragments at the right size. We will do the ligation again.
 +
|}
 +
{| border="1" align="center"
{| border="1" align="center"
Line 75: Line 145:
|[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]]
|[[Team:Paris_Saclay/Notebook|<big>Back to calendar</big>]]
-
|[[Team:Paris Saclay/Notebook/July/17|<big>Next day</big>]]
+
|[[Team:Paris Saclay/Notebook/July/17|<big>next day</big>]]
|}
|}
{{Team:Paris_Saclay/incl_fin}}
{{Team:Paris_Saclay/incl_fin}}

Revision as of 19:11, 26 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : July 16

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003, Bba_K1155007

1 - Extraction of Bba_B0015, Bba_B0017, Bba_I732019 from DH5α

Zhou

Protocol : High copy plamid extraction

2 - Digestion of Bba_B0015, Bba_B0017, Bba_I732019 by EcoRI/PstI

Anaïs

  • DNA : 5µL
  • Buffer FD : 2µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • H2O : 21µL

We let our digestion 1h30 at 37°C.

Objective : obtaining biobricks in PSB3K3

1 - Digestion of PSB3K3 by EcoRI/PstI

Anaïs

Used quantities :

  • DNA : 5µL
  • Buffer FD : 2µL
  • EcoRI FD : 1µL
  • PstI FD : 1µL
  • H2O : 11µL

We let our digestion 1h30 at 37°C.

2 - Electrophoresis of the digestion of PSB3K3 by EcoRI/PstI

Sheng

[[]]
  • Well 1 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 2 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 3 : 6µL of DNA ladder
  • Well 4 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Well 5 : 5µL of PSB3K3 digested by EcoRI/PstI+1µL of 6X loading dye
  • Gel : 1%

Expected sizes :

  • PSB3K3 : 2750bp

We obtain fragments at the right size.

3 - Gel purification of electrophoresis of the digestion of PSB3K3 by EcoRI/PstI

Abdou

Protocol : [http://www.mn-net.com/tabid/1452/default.aspx Gel purification ]

B - PCB sensor system

Objective : obtaining Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3

1 - Electrophoresis of the digestion of Bba_K1155001, Bba_K1155002, BphR2 in PSB1C3

Anaïs, Zhou

  • Bba_K1155001 :
[[]]
  • Well 1 to 5 : 2µL Bba_K1155001 digested by EcoRI/PstI+2µl of 6X loading dye
  • Well 6 and 7 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
  • Well 8 : 6µL DNA Ladder
  • Gel : 1.2%

Expected size :

  • Bba_K1155001 digested by EcoRI/PstI : 2037bp + 333bp
  • Bba_K1155001 digested by SacII : 2370bp

In the first well, we obtain fragment at the right size. We will amplify Bba_K1155001.

  • Bba_K1155002 :
[[]]
  • Well 1 to 8 : 2µL Bba_K1155002 digested by EcoRI/PstI+2µl of 6X loading dye
  • Well 9 : 6µL DNA Ladder
  • Well 10 to 17 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
  • Gel : 1.5%

Expected size :

  • Bba_K1155002 digested by EcoRI/PstI : ...
  • Bba_K1155002 digested by SacII : ...

We didn't obtain fragments at the right size. We will do the ligation again.

  • BphR2 in PSB1C3 :
[[]]
  • Well 1 to 8 : 2µL Bba_K1155002 digested by EcoRI/PstI+2µl of 6X loading dye
  • Well 9 : 6µL DNA Ladder
  • Well 10 to 17 : 2µL Bba_K1155001 digested by SacII+2µl of 6X loading dye
  • Gel : 1.5%

Expected size :

  • Bba_K1155002 digested by EcoRI/PstI : ...
  • Bba_K1155002 digested by SacII : ...

We didn't obtain fragments at the right size. We will do the ligation again.


Previous day Back to calendar next day

Retrieved from "http://2013.igem.org/Team:Paris_Saclay/Notebook/July/16"