"
Team:Evry/Experimentals Parameters
From 2012.igem.org
PierreYves (Talk | contribs) |
PierreYves (Talk | contribs) |
||
| Line 12: | Line 12: | ||
<br/> | <br/> | ||
| - | <center><h2> | + | <center><h2>Identify where the auxin concentrates</h2></center> |
<br/> | <br/> | ||
| + | <br/> | ||
| + | |||
| + | <p> To determine where the auxin concentrates, you have to inject first the auxin in the heart of the tadpole, for it is directly associated into the bloodstream. Then, after waiting 20 minutes that auxin diffuses throughout the body, fixing the tadpole to permeabilize the tissue, then a revelation by fluorescent antibody anti-auxin, visualizes the areas where concentrated auxin. </p> | ||
| + | |||
| + | |||
<br/> | <br/> | ||
<br/> | <br/> | ||
| - | <center> | + | <center><h2>How to determinate the Auxin diffusion from Skin to Water</h2></center> |
| - | < | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | </center> | + | |
<br/> | <br/> | ||
| + | <br/> | ||
| + | |||
| + | <p> To determine the diffusion coefficient of auxin from the skin to the environment (MMR), just in time to first soak for 20 minutes a tadpole skin volume, the volume has been previously measured in a solution MMR with auxin. Then transferring the tissue in a solution of MMR, and every 5 minutes taking a constant volume of media for each samples it is possible to determine the desired parameter. In fact, each samples shows the quantity of auxin diffused from the skin to the media, during a period. It is thus possible to plot a curve ( quantities fonction of time ), and determine the parameters about Auxin diffusion from Skin to Water. </p> | ||
| + | |||
| + | |||
<br/> | <br/> | ||
<br/> | <br/> | ||
| Line 31: | Line 36: | ||
<center><h2>How to determinate the Auxin diffusion from Blood to Muscle</h2></center> | <center><h2>How to determinate the Auxin diffusion from Blood to Muscle</h2></center> | ||
| - | |||
<br/> | <br/> | ||
<br/> | <br/> | ||
| - | < | + | <p> To determine the diffusion coefficient of auxin, it will have injected directly into the heart auxin. Then let the tadpole in the MMR for 5 minutes ( recuperation time ). |
| - | + | Then euthanize the tadpole and collect a volume of muscle. | |
| - | + | Determining the concentration of auxin in the piece of muscle removed. | |
| - | + | Repeat the experiment with at least 10 others tadpoles increasing the recuperation time of 5 minutes by experiments. | |
| - | + | Remark, you have to isolated always the same height of muscle. | |
| - | + | Thus, the values of auxin diffused from the blood to the muscle function of time will determine the desired parameters. </p> | |
| + | |||
| - | |||
<br/> | <br/> | ||
<br/> | <br/> | ||
| Line 52: | Line 56: | ||
<br/> | <br/> | ||
| - | < | + | <p> To determine the diffusion coefficient of auxin, it will have injected directly into the heart auxin. Then let the tadpole in the MMR for 5 minutes ( recuperation time ). |
| - | + | Then euthanize the tadpole and collect a volume of muscle. | |
| - | + | Determining the concentration of auxin in the piece of muscle removed. | |
| - | + | Repeat the experiment with at least 10 others tadpoles increasing the recuperation time of 5 minutes by experiments. | |
| - | + | Remark, you have to isolated always the same height of muscle. | |
| - | + | Thus, the values of auxin diffused from the blood to the muscle function of time will determine the desired parameters. </p> | |
| + | |||
<br/> | <br/> | ||
Revision as of 23:33, 26 September 2012
Protocols to determinate the following experimentals parameters
Identify where the auxin concentrates
To determine where the auxin concentrates, you have to inject first the auxin in the heart of the tadpole, for it is directly associated into the bloodstream. Then, after waiting 20 minutes that auxin diffuses throughout the body, fixing the tadpole to permeabilize the tissue, then a revelation by fluorescent antibody anti-auxin, visualizes the areas where concentrated auxin.
How to determinate the Auxin diffusion from Skin to Water
To determine the diffusion coefficient of auxin from the skin to the environment (MMR), just in time to first soak for 20 minutes a tadpole skin volume, the volume has been previously measured in a solution MMR with auxin. Then transferring the tissue in a solution of MMR, and every 5 minutes taking a constant volume of media for each samples it is possible to determine the desired parameter. In fact, each samples shows the quantity of auxin diffused from the skin to the media, during a period. It is thus possible to plot a curve ( quantities fonction of time ), and determine the parameters about Auxin diffusion from Skin to Water.
How to determinate the Auxin diffusion from Blood to Muscle
To determine the diffusion coefficient of auxin, it will have injected directly into the heart auxin. Then let the tadpole in the MMR for 5 minutes ( recuperation time ). Then euthanize the tadpole and collect a volume of muscle. Determining the concentration of auxin in the piece of muscle removed. Repeat the experiment with at least 10 others tadpoles increasing the recuperation time of 5 minutes by experiments. Remark, you have to isolated always the same height of muscle. Thus, the values of auxin diffused from the blood to the muscle function of time will determine the desired parameters.
How to determinate the Auxin Half Life in the Skin
To determine the diffusion coefficient of auxin, it will have injected directly into the heart auxin. Then let the tadpole in the MMR for 5 minutes ( recuperation time ). Then euthanize the tadpole and collect a volume of muscle. Determining the concentration of auxin in the piece of muscle removed. Repeat the experiment with at least 10 others tadpoles increasing the recuperation time of 5 minutes by experiments. Remark, you have to isolated always the same height of muscle. Thus, the values of auxin diffused from the blood to the muscle function of time will determine the desired parameters.
