"
Team:Groningen/16 August 2010
From 2010.igem.org
| Line 2: | Line 2: | ||
'''Week 25''' | '''Week 25''' | ||
| + | |||
<br> | <br> | ||
| - | + | ||
| + | ''David'' | ||
| + | |||
<br> | <br> | ||
| - | + | ||
| + | '''THT Staining - Chaplin ladder''' | ||
| + | |||
<br> | <br> | ||
| + | |||
| + | '''THT reference'' | ||
| + | |||
| + | Making a reference of pure chaplins is of great importance when comparing results. For a reference 22 mg of extracted and freeze-dried celwalls of streptomyces were used. Chaplins from these cellwalls were then purified by TFA treatment. The monomerized chaplins were diluted in 1 ml demiwater. Then a variety of dilutions was made with demiwater. | ||
| + | |||
| + | Amount of diluted chaplin protein in a 250 ul sample (high concentration reference): | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <pre> | ||
| + | 100% | ||
| + | 51,2% | ||
| + | 25,6% | ||
| + | 12,8% | ||
| + | 6,4% | ||
| + | 3,2% | ||
| + | 1,6% | ||
| + | 0,8% | ||
| + | 0,4% | ||
| + | |||
| + | Blanco(demi+THT staining) | ||
| + | |||
| + | demi water | ||
| + | </pre> | ||
| + | |||
| + | <br> | ||
| + | |||
| + | <pre> | ||
| + | Low concentration ladder: | ||
| + | 0,8% | ||
| + | 0,6% | ||
| + | 0,4% | ||
| + | 0,3% | ||
| + | 0,2% | ||
| + | blanco | ||
| + | demi | ||
| + | </pre> | ||
| + | |||
| + | <br> | ||
| + | |||
'''Expression experiment''' - ''David & Peter'' | '''Expression experiment''' - ''David & Peter'' | ||
| + | |||
<br> | <br> | ||
| + | |||
The first expression experiment, testing all the constructs that were perhaps made so far. Tested for pellet and supernatant, raw pellets were tested as well. | The first expression experiment, testing all the constructs that were perhaps made so far. Tested for pellet and supernatant, raw pellets were tested as well. | ||
<br> | <br> | ||
Revision as of 15:45, 24 October 2010
Hydrophobofilm
pushing coatings into a greener future
Week 25
David
THT Staining - Chaplin ladder
'THT reference
Making a reference of pure chaplins is of great importance when comparing results. For a reference 22 mg of extracted and freeze-dried celwalls of streptomyces were used. Chaplins from these cellwalls were then purified by TFA treatment. The monomerized chaplins were diluted in 1 ml demiwater. Then a variety of dilutions was made with demiwater.
Amount of diluted chaplin protein in a 250 ul sample (high concentration reference):
100% 51,2% 25,6% 12,8% 6,4% 3,2% 1,6% 0,8% 0,4% Blanco(demi+THT staining) demi water
Low concentration ladder: 0,8% 0,6% 0,4% 0,3% 0,2% blanco demi
Expression experiment - David & Peter
The first expression experiment, testing all the constructs that were perhaps made so far. Tested for pellet and supernatant, raw pellets were tested as well.
Analasys was done by THT and SDS page.
Modellers: More literature research on ComXPA. Joël, Laura. Researching parts registry for information which is required to build an information standard Arend
