Team:Freiburg/Notebook/26 July

From 2011.igem.org

(Difference between revisions)
(→Precipitator)
(→Precipitator)
Line 144: Line 144:
3A assembly combination as shown in table.
3A assembly combination as shown in table.
 +
 +
|}
 +
 +
 +
Transformation''' '''
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name: RT
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 26.07
 +
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Date Name
 +
 +
Experiment Ligation 26.07 Ruediger
 +
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name: GFPPbd
 +
 +
|}
 +
Procedure
 +
 +
 +
# take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells)
 +
# thaw cells on ice 20 minutes
 +
# pipette 50 μl cells and 2 μl DNA into eppi still on ice!
 +
# Incubate for 30 minutes on ice
 +
# Heat at 42°C for 60 sec
 +
# Incubate on ice for 5 minutes
 +
# Add 200 μl LB Broth
 +
# Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!)
 +
# Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance
 +
 +
'''Documentation:'''
 +
 +
Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc.
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Had only 6 tet plates -> put 100 μl of each sample on each plate
 +
 +
Incubator overnight for 30°C
 +
 +
 +
S39-P18
 +
 +
S39-P19
 +
 +
S39-P20
 +
 +
 +
S43-P18
 +
 +
S43-P19
 +
 +
S43-P20
|}
|}

Revision as of 09:35, 5 August 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

Gibson-Assembly

Investigators: Sandra, Sophie

Planned Gibson-Assembly for Lov-tap and Not-Gate. Need to order NAD. We will start an thursday.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Ligation


Name: Ruediger Date: 26.07
Continue from Date Name

Experiment Digestion 25.07 Ruediger

Project Name:GFPPbd

Procedure


PCR tube:

total volume 20 μl


  1. add H2O (17 μl -X-Y-Z)
  2. add 2 μl Ligase Buffer 10x
  3. add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
  4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
  5. Add 1 μl T4-DNA Ligase
  6. Incubate 10-30 min at room temperature
  7. heat for 20 minutes at 80°C
  8. store at -20°C or directly proceed to transformation


Part name Volume (μl) Part name Volume (μl)
S39 2 S43 2
P18 1,8 P18 1,8
Psb1T3 2 Psb1T3 2
S39 2 S43 2
P19 1,8 P19 1,8
Psb1T3 2 Psb1T3 2
S39 2 S43 2
P20 1,8 P20 1,8
Psb1T3 2 Psb1T3 2

Added 12,2 μl to every sample


Documentation:

Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.


See digestion yesterday

3A assembly combination as shown in table.


Transformation


Name: RT Date: 26.07
Continue from Date Name

Experiment Ligation 26.07 Ruediger

Project Name: GFPPbd

Procedure


  1. take cells from -80°C freezer and put them on ice! (every eppi contains about 400 μl cells)
  2. thaw cells on ice 20 minutes
  3. pipette 50 μl cells and 2 μl DNA into eppi still on ice!
  4. Incubate for 30 minutes on ice
  5. Heat at 42°C for 60 sec
  6. Incubate on ice for 5 minutes
  7. Add 200 μl LB Broth
  8. Incubate for 2 hours at 37°C (cells with lysis cassette at 30°C!!)
  9. Plate 50 μl and 200μl on two different LB/Agar plates with appropriate antibiotic resistance

Documentation:

Why are you doing this experiment? Name of the sample? Where are they stored? Name the vector with inserts, antibiotika resistance etc.


Had only 6 tet plates -> put 100 μl of each sample on each plate

Incubator overnight for 30°C


S39-P18

S39-P19

S39-P20


S43-P18

S43-P19

S43-P20

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/26_July"