Team:Paris Saclay/Notebook/July/23

From 2013.igem.org

(Difference between revisions)
Line 53: Line 53:
{|
{|
-
| style="width:350px;border:1px solid black;" |[[File:Psgel32307.jpg|300px]]
+
| style="width:350px;border:1px solid black;" |[[File:Psgel32307.jpg|200px]]
| style="width:350px;border:1px solid black;vertical-align:top;" |
| style="width:350px;border:1px solid black;vertical-align:top;" |
* Well 1 : 6µL of DNA 6X loading dye
* Well 1 : 6µL of DNA 6X loading dye

Revision as of 00:17, 29 September 2013

Navigation Home Team Project Labwork Notebook Human practices

Contents

Notebook : July 23

Lab work

A - Aerobic/Anaerobic regulation system

Objective : obtaining Bba_K1155003, Bba_K1155007

1 - Electrophoresis of the digestion of Bba_I732017 by EcoRI/SpeI, Bba_K592009 by SpeI/PstI, Bba_B0015, Bba_B0017 by XbaI/PstI

Abdou, Anaïs, Xavier, Zhou

Psgel12307.jpg
  • Well 1 to 3: 30µL of Bba_I732017 digested by EcoRI/SpeI+6µL of 6X loading dye
  • Well 4 : 6µL of DNA 6X loading dye
  • Gel : 0.8%
Psgel22307.jpg
  • Well 1 : 5µL of Bba_B0015 digested by XbaI/PstI+1µL of 6X loading dye
  • Well 2 : 5µL of Bba_B0017 digested by XbaI/PstI+1µL of 6X loading dye
  • Well 3 : 5µL of Bba_K592009 digested SpeI/PstI+1µL of 6X loading dye
  • Well 4 : 6µL of DNA 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ : 3093bp
  • Amil CP :
  • Term :
  • PSB3A2 : 2079bp

We obtain a RBS-lacZ and Term fragments at the right size. The digestions were good and we will purify them.

2 -Gel purification of digestion products : RBS-LacZ

Xavier

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

3 - Electrophoresis to check the gel purification of digestion products : RBS-LacZ

Abdou

Psgel32307.jpg
  • Well 1 : 6µL of DNA 6X loading dye
  • Well 2: 5µL of RBS-LacZ+1µL of 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ :

We can't see anything. The purification wasn't good. We will do the digestion again.

4 - Digestion of Bba_I732017 by EcoRI/PstI

Anaïs

Used quantities :

  • Bba_I732017 : 10µL
  • Buffer FD : 3µL
  • EcoRI FD : 1.5µL
  • PstI FD : 1.5µL
  • H2O : 14µL

5 - Electrophoresis to check the gel digestion of Bba_I732017 by EcoRI/SpeI

Zhou

Psgel42307.jpg
  • Well 1 : 6µL of DNA 6X loading dye
  • Well 2 to 4 : 5µL of Bba_I732017 digested by EcoRI/SpeI+1µL of 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • RBS-LacZ :

We can't see anything. The digestion wasn't good, we will do it the again.

6 - Digestion of Bba_K592009 by SpeI/PstI

Anaïs

Used quantities :

  • Buffer FD : 3µL
  • DNA : 15µL
  • SpeI FD : 2µL
  • PstI : 2µL
  • H2O : 8µL

7 - Electrophoresis to check the digestion of Bba_K592009 by SpeI/PstI

Zhou

[[]]
  • Well 1 : 6µL of DNA 6X loading dye
  • Well 2 to 4 : 5µL of Bba_K592009 digested by SpeI/PstI+1µL of 6X loading dye
  • Gel : 0.8%

Expected sizes :

  • Amil CP :

We can't see anything. The purification wasn't good. We will do cultures again.

8 - Liquid culture of Bba_I732017 and Bba_K592009 in DH5α

Abdou, Xavier

We let our preparations at 37°C, 150rpm with 5mL of LB and 5mL of chloramphenicol antibiotic.

Previous day Back to calendar Next day

Retrieved from "http://2013.igem.org/Team:Paris_Saclay/Notebook/July/23"