Team:Freiburg/Notebook/17 August

From 2011.igem.org

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(→Testdigest)
(→blue light receptor)
Line 109: Line 109:
*EcoRI
*EcoRI
*PstI
*PstI
 +
===3A-assembly of Not-Gate, LovTAP in pSB1T3===
 +
 +
'''Investigators: Sophie'''
 +
 +
'''Digestion'''
 +
 +
Amount of DNA and H20:
 +
 +
{| cellpadding="10" cellspacing="0" border="1"
 +
|Sample
 +
|DNA μl
 +
|H20 μl
 +
|-
 +
|LovTAP
 +
|5
 +
|33
 +
|-
 +
|Not-Gate
 +
|2
 +
|36
 +
|-
 +
|pSB1T3
 +
|20
 +
|18
 +
|}
 +
 +
 +
Enzymes necessary for digestion:
 +
 +
{|cellpadding="10" cellspacing="0" border="1"
 +
|
 +
|LovTAP
 +
|Not-Gate
 +
|vector
 +
|-
 +
|enzyme 1
 +
|EcoRI
 +
|XbaI
 +
|EcoRI
 +
|-
 +
|enzyme 2
 +
|NheI
 +
|PstI
 +
|PstI
 +
|}
 +
 +
 +
*Incubation at 37°C for 6 hours
 +
*20 minutes at 80°C
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==

Revision as of 13:42, 17 August 2011


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

Contents

Commons

PCR


Name: Sophie


Date: 17.08.11
Continue from Experiment (Date)

(Name): Commons

Project Name: tet-vector seems to make problems -> new PCR with original psB1T3-DNA

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 SB-prep-3P-1
2.5µl Primer dw SB-prep-2Ea
1µl dNTPs of Template DNA
1µl DNA-Template

PSB 1 T3

0.5 µl Phusion (add in the end)

What program do you use?

  1. 2Min 94°C
  2. 30s 94°C
  3. 30s 55°C
  4. 3min 72°C
  5. 10min 72°C

step 2,3 and 4 in 35 cycles


To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.


How did you label the PCR-Product, where is it stored and what do you do next?

Labeled PSB 1 T3 stored in -20°C in last drawer

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

blue light receptor

Miniprep

Investigators: Sandra

Miniprep of LovTAP-NotGate-T3:

  • 15ng/μl

stored in undigested miniprep box II


Testdigest

Investigators: Sandra

Testdigest of Miniprep product.

Enzymes:

  • EcoRI
  • PstI

3A-assembly of Not-Gate, LovTAP in pSB1T3

Investigators: Sophie

Digestion

Amount of DNA and H20:

Sample DNA μl H20 μl
LovTAP 5 33
Not-Gate 2 36
pSB1T3 20 18


Enzymes necessary for digestion:

LovTAP Not-Gate vector
enzyme 1 EcoRI XbaI EcoRI
enzyme 2 NheI PstI PstI


  • Incubation at 37°C for 6 hours
  • 20 minutes at 80°C

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

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