Promocionado por PROMEGA NL
Universidad Catolica de Louvain, Escuela de medicine (Bruselas, Belgium)


atelier de genetica molecular SBIM 2520
disección
regresa a la primera semana de trabajo

Maritza BARCIA MACAY* (Pharmacist), Daniel RUIZ** (Erasmus), Jean-Pierre HERVEG (Emeritus)***
*Universidad de Guayaquil, Ecuador, ** Universidad de Alcalá, Madrid, España, ***Universidad Catolica de Louvain, Belgica


PLAN

1. prepare he homogenizer
2.
tools

3. needle the mouse
4.
cut the skin
5. cut off the
salivary glands

6. dissociate muscles
7. discover the
trachea
8. dissociate
lateral muscles
9. discover the thyroid
10. dissect and
homogenize the gland
11. extract
total RNA (on another html document)



In the Promega kit, SV Total RNA Isolation, take the brown bottle containing the RNA lysis buffer.
Prepare a Potter homogenizer treated against RNAse.

Prepare an container with ice and water.

Place the homogenizer inside this cooling system, and remove the pestle.

add 200 µl ß-mercapto-ethanol (BME) to 10 ml of Lysis Buffer. Store at 4°C. Note on the bottle that BME was added...!


Pipet 170 µl of SV RNA lysis buffer into the homogenizer. Put the pestle back. Now, kill the mouse with Chloroform and start dissection.

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1. herramientas

ajustar (calibrar),
el brazo del microscopio
cambio de objetivos
plato de dissecci
Ón
elecci
Ón del interruptor correcto



2. pinchar el ratÓn

regresa al comienzo


tigeras grandes
pinzas grandes (sin dientes)
agujas inclinadas


3. haz una incisiÓn en la piel






regresa al comienzo


4. eliminaciÓn de las glándulas salivares

5. liberar los músculos

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6. liberar la traquea

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7. corta los musculos laterales

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8. reveal the thyroid

slide a needle underneath the trachea. It makes things a lot easier to find and dissect the whole gland.

On the picture on your left, we inserted a needle through the trachea to help showing the two thyroid lobes.

 

 

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9. dissect, homogenize each lobe quickly


Dissect the first lobe and put it on the pestle as shown to your right. Homogenize it directly at 0° C

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extract total RNA

Promocionado por PROMEGA NL
Universidad Catolica de Louvain, Escuela de medicine (Bruselas, Belgium)


atelier de genetica molecular SBIM 2520
disección
regresa a la primera semana de trabajo

Maritza BARCIA MACAY* (Pharmacist), Daniel RUIZ** (Erasmus), Jean-Pierre HERVEG (Emeritus)***
*Universidad de Guayaquil, Ecuador, ** Universidad de Alcalá, Madrid, España, ***Universidad Catolica de Louvain, Belgica