Team:Freiburg/Notebook/9 September

From 2011.igem.org

(Difference between revisions)
(→blue light receptor)
(→blue light receptor)
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==<span style="color:blue;">blue light receptor</span>==
==<span style="color:blue;">blue light receptor</span>==
-
===Transformatoin===
+
'''Digestion'''
-
'''Investigators:Sophie'''
 
-
Transformation with the Gibson-♥-NOT + PR-Vector Ligation
+
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name: Sophie
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 9.9.11
 +
 
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Date 9.9.11 Name: Sophie
 +
 
 +
Experiment: PCR
 +
 
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name: ♥-NOT in PR-vector
 +
 
 +
|}
 +
Procedure
 +
 
 +
 
 +
# add H<sub>2</sub>O (38μl-DNA )
 +
# 5 μl NEB4 buffer  (stored at iGEM’s, -20°C)
 +
# 5 μl 10x BSA  (used 1:10 diluted sample stored at iGEM’s, -20°C)
 +
# DNA (500 ng) (Vector:Insert ratio 1:3 in following Ligation)
 +
# 1 μl restriction enzymes  (stored at iGEM’s, -20°C)
 +
# heat for 1-2 hours 37°C (6 hours if time)
 +
# heat for 20 minutes 80°C (inactivation of enzymes)
 +
# keep at 4°C if you cannot continue
 +
 
 +
Measured DNA-concentration with Nanodrop to calculate the volume of DNA to do the digestion:
 +
 
 +
 
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Sample Name
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA concentration (μg/μl)
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| G-♥-NOT a,b
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| ~140
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 
 +
|}
 +
Restriction enzymes you need to cut the vector, insert1 and insert 2:
 +
 
 +
 
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| '''Components'''
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| <center>'''Vector (μl)'''</center>
 +
| colspan="2"  style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| '''Insert(μl)'''
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA (500ng)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| BSA (10x) (5μl)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| NEB4 Buffer (5μl)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Enzyme 1 (1μl)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Spe I
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Nhe I
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Enzyme 2 (1μl)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Pst I
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Pst I
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>O (38 μl- DNA)
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
||
 +
 
 +
|-
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| In total 50 μl
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
||
 +
 
 +
|}
 +
'''Documentation:'''
 +
 
 +
Why are you doing this experiment? Where are the samples stored? Antibiotica resistance, vector used etc.
 +
 
 +
 
 +
 
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #000000;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| The LOV-Repressor protein needs to be expressible
 +
 
 +
Samples stored in blue light box
 +
 
 +
Vector used: all PR-vectors (D39- D44)
 +
 
 +
|}
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==

Revision as of 12:09, 10 September 2011


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

Contents

Commons

Testdigests

Investigators: Sophie

Testdigest of our various minipreps

100Testdigests Freiburg.jpg


green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Digestion


Name: Sophie Date: 9.9.11
Continue from Date 9.9.11 Name: Sophie

Experiment: PCR

Project Name: ♥-NOT in PR-vector

Procedure


  1. add H2O (38μl-DNA )
  2. 5 μl NEB4 buffer (stored at iGEM’s, -20°C)
  3. 5 μl 10x BSA (used 1:10 diluted sample stored at iGEM’s, -20°C)
  4. DNA (500 ng) (Vector:Insert ratio 1:3 in following Ligation)
  5. 1 μl restriction enzymes (stored at iGEM’s, -20°C)
  6. heat for 1-2 hours 37°C (6 hours if time)
  7. heat for 20 minutes 80°C (inactivation of enzymes)
  8. keep at 4°C if you cannot continue

Measured DNA-concentration with Nanodrop to calculate the volume of DNA to do the digestion:


Sample Name DNA concentration (μg/μl)
G-♥-NOT a,b ~140

Restriction enzymes you need to cut the vector, insert1 and insert 2:


Components
Vector (μl)
 Insert(μl)
DNA (500ng)
BSA (10x) (5μl)
NEB4 Buffer (5μl)
Enzyme 1 (1μl) Spe I Nhe I
Enzyme 2 (1μl) Pst I Pst I
H2O (38 μl- DNA)
In total 50 μl

Documentation:

Why are you doing this experiment? Where are the samples stored? Antibiotica resistance, vector used etc.


The LOV-Repressor protein needs to be expressible

Samples stored in blue light box

Vector used: all PR-vectors (D39- D44)

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/9_September"