Team:Freiburg/Notebook/16 August

From 2011.igem.org

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==<span style="color:orange;">Lysis cassette</span>==
==<span style="color:orange;">Lysis cassette</span>==
-
===NAME OF YOUR EXPERIMENT===
+
===2A Assembly like August 15===
-
'''Investigators:NAME'''
+
'''Investigators:Theo'''
 +
This time I used all the amount of S15 plasmid DNA we had, did an overnight restriction and run the gel.<br>
 +
[[File:Freiburg11 1608gelextraction.png]]
==<span style="color:grey;">Precipitator</span>==
==<span style="color:grey;">Precipitator</span>==

Revision as of 23:10, 21 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Picking clones

Investigators: Sandra

Picking clones of transformed cells with LovTAp, NotGate, pSB1T3.

red light receptor

continuing with PCR of cph8

Investigators:Julia
2µl of the PCR reaction from 15th Aug. were loaded onto a gel.
But there was no band. Jakob is going to repeat the PCR.

3a-assembly with promotor+ribosome binding site infront of pcyA and ho1

Investigators:Julia

Digestion

  • PR1 in pSB1C3
  • PR2 in pSB1C3
  • pcyA (pSB1T3)
  1. pcyA+terminator(d)in pSB1T3
  • ho1+Terminator (pSB1T3)
  • vector (pSB1K3)


Amount of DNA and H20:

Sample 500(ng)/DNA concentration (ng/μl) H20 μl
pcyATd 2.8 35.2
pcyaTb 2.9 35.1
PR2 7.2 30.8
PR1 6.1 31.9
S22a 2,5 35.5
pSB1K3 20 18


Enzymes necessary for digestion:

PR1/PR2 pcyA/S22a vector
enzyme 1 EcoRI XbaI EcoRI
enzyme 2 SpeI PstI PstI


-incubated for 1 hours at 37°C.
-heated for 20 minutes at 80°C


Ligation

Ligation of:

PcyATd S22a (ho1)
PR1 1 pTd 1 pTb
PR2 2 pTd 2 pTb

PCR

Investigator: Jakob


PCR


Name:

Jakob

Date:

16.08.2011

Continue from Experiment: 12.08.2011

Order primers

Project Name:

Red light receptor, amplify Cph8

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20 Name
10µl 5x Phusion Buffer of Primer
2.5µl Primer fw P 58 (1:10)
2.5µl Primer dw P 59 (1:10)
1µl dNTPs of Template DNA
1µl DNA-Template JT122 5ng/µl
0.5 µl Phusion (add in the end)

The program we used:


Temp.
Time
 Nr. of cycles
98°
5´
1x
98°
30´´
10x
65°
30´´
72°
1´5´´
98°
30´´
20x
72°
1´5´´
72°
7´
1x
4°
∞


We got the information for the program and the primer sequences from the iGEM Team Uppsala Sweden.

Lysis cassette

2A Assembly like August 15

Investigators:Theo


This time I used all the amount of S15 plasmid DNA we had, did an overnight restriction and run the gel.
Freiburg11 1608gelextraction.png

Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

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