Today, accompanied Miss. Vinita (JRF) while she was working on SDS- PAGE for analysing protein samples. Learnt practical concepts of preparing gels ( stacking and resolving gels) and running buffer for the running of protein samples.
Then the gel was analyzed using Coomassie brilliant blue dye. The steps include:
1.Fixation
2. Staining
3. Destaining ( Protocol )
While doing the above practical, learnt the importance of using different pH in stacking and resolving gel. ( know more ) and also learnt to maintain pH.
To measure pH, we had to use PTC lab as the pH meter was available only in that lab. The infrastructure of the lab was very well designed to keep it sterile. At the entrance, there is a entry gate and a window beside it. All the material (glassware, pipettes, or any instrument or objects) that are supposed to be taken into the lab should be deposited in the window (it has its second opening in the interior of the Lab from where things have to be collected.). Foot ware had to be placed in the shoe rack beside the entrance. Once we enter the "ENTRY" there is a closed chamber with air blower in it. It senses our presence and the blowers are switched on. In the same chamber, the workers have to wear the footware specially kept for the PTC lab purpose. After few seconds, another door opens from the air shower chamber into the interior of the PTC lab. Here, general instruments such as centrifuge machine, pH meter, Magnetic Stirrer and other chemicals were kept. Other isolated labs are present in separated chambers where laminar airflow cabinets were placed and workers worked under extremely sterile conditions.
It was a long day and learnt too many things. A good day afterall.
Good night.
Then the gel was analyzed using Coomassie brilliant blue dye. The steps include:
1.Fixation
2. Staining
3. Destaining ( Protocol )
While doing the above practical, learnt the importance of using different pH in stacking and resolving gel. ( know more ) and also learnt to maintain pH.
To measure pH, we had to use PTC lab as the pH meter was available only in that lab. The infrastructure of the lab was very well designed to keep it sterile. At the entrance, there is a entry gate and a window beside it. All the material (glassware, pipettes, or any instrument or objects) that are supposed to be taken into the lab should be deposited in the window (it has its second opening in the interior of the Lab from where things have to be collected.). Foot ware had to be placed in the shoe rack beside the entrance. Once we enter the "ENTRY" there is a closed chamber with air blower in it. It senses our presence and the blowers are switched on. In the same chamber, the workers have to wear the footware specially kept for the PTC lab purpose. After few seconds, another door opens from the air shower chamber into the interior of the PTC lab. Here, general instruments such as centrifuge machine, pH meter, Magnetic Stirrer and other chemicals were kept. Other isolated labs are present in separated chambers where laminar airflow cabinets were placed and workers worked under extremely sterile conditions.
It was a long day and learnt too many things. A good day afterall.
Good night.
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