Difference between revisions of "Team:elan vital korea/Protocol"
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− | 1. Reagents used in our project, such as restriction reagents, must be stored in low temperature. The reagents must be stored in the freezer when they are not used, and must be put on ice when taking them out of the freezer for an experiment. | + | <b>1.</b> |
+ | Reagents used in our project, such as restriction reagents, must be stored in low temperature. The reagents must be stored in the freezer when they are not used, and must be put on ice when taking them out of the freezer for an experiment. <br><br> | ||
− | 2. Reagents should be added last to the solution, because reagents are sensitive to inactivation by pH and ionic conditions that deviate from their storage and reaction buffers. After adding reagents, the mixed solution should be mixed completely. | + | <b>2.</b> |
+ | Reagents should be added last to the solution, because reagents are sensitive to inactivation by pH and ionic conditions that deviate from their storage and reaction buffers. After adding reagents, the mixed solution should be mixed completely. | ||
</font> | </font> | ||
</p> | </p> | ||
</div> | </div> | ||
+ | <h5 style="text-align:center;"> | ||
+ | <font color="black"> | ||
+ | Protocols to store materials and maintain <Br> | ||
+ | usage history of each material. | ||
+ | </font> | ||
+ | </h5> | ||
+ | |||
+ | <div class="inner0"> | ||
+ | <P style="text-align:left;"> | ||
+ | <font color="black"> | ||
+ | <b>1.</b> | ||
+ | Reporter cell, test cell and competent cell (Top 10 invitrogen) must be kept at 4°C and frequently used reagents, reagents, DNA plasmids should be kept at −20°C in the freezer. <br><br> | ||
+ | |||
+ | <b>2.</b> | ||
+ | We use triple distilled water (or DDH2O) to make LB broth. Triple distilled water is kept at lab temperature (around 18 °C or lower). <br><br> | ||
+ | |||
+ | <b>3.</b> | ||
+ | Other materials such as yeast and NaCl are stored and maintained under the responsibility of Gachon Molecular Biology Lab. <br><br> | ||
+ | |||
+ | <b>4.</b> | ||
+ | We have to record the history of each material, including if plasmids/reporter cell/ test cell/ AHL have been frozen and if so, when it is used. <br><br> | ||
+ | </font> | ||
+ | </p> | ||
+ | </div> | ||
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− | + | LB Medium | |
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Revision as of 19:46, 17 September 2015
Protocols
We conducted our experiments by following the protocols below. As an official procedure, lab workers should understand the lab experiment assigned to them along with safety procedures before starting lab work. The protocols are arranged according to the order of experiments we followed.
How to handle reagents.
1.
Reagents used in our project, such as restriction reagents, must be stored in low temperature. The reagents must be stored in the freezer when they are not used, and must be put on ice when taking them out of the freezer for an experiment.
2.
Reagents should be added last to the solution, because reagents are sensitive to inactivation by pH and ionic conditions that deviate from their storage and reaction buffers. After adding reagents, the mixed solution should be mixed completely.
Protocols to store materials and maintain
usage history of each material.
1.
Reporter cell, test cell and competent cell (Top 10 invitrogen) must be kept at 4°C and frequently used reagents, reagents, DNA plasmids should be kept at −20°C in the freezer.
2.
We use triple distilled water (or DDH2O) to make LB broth. Triple distilled water is kept at lab temperature (around 18 °C or lower).
3.
Other materials such as yeast and NaCl are stored and maintained under the responsibility of Gachon Molecular Biology Lab.
4.
We have to record the history of each material, including if plasmids/reporter cell/ test cell/ AHL have been frozen and if so, when it is used.
To The Top
LB Medium
Other significant Safety Considerations.
1. Fire Regulations.
When fire breaks out, the following guidelines shall apply.
Also it is very important to know the locations of the fire-fighting equipment, fire alarms, and evacuation routes closest to the lab.
A small fire can be extinguished quickly by smothering it in a fire blanket or by spraying it with a fire extinguisher.
Lab fires in biological labs are caused most commonly by the plating of cell cultures.
To prevent this,
CHEMICALS
On each chemical container, there is a label that specifies the potential danger of the substance for humans and/or the environment.
Chemicals should be handled cautiously with gloves, both for your safety and for decreasing the contamination risk.
Always wear a lab coat and shoes as additional protection. Read the signs on the chemical container and the Material Safety Data Sheet
(MSDS; available online) for further direction.
BIO- SAFETY AND DISPOSAL
When you are working to generate new organisms by molecular methods, you are regulated in most countries
according to international biosafety guidelines:
When working with microorganisms such as bacteria and viruses, there are four BioSafety Levels (BSL) numbered BSL1–4.
Biosafety level is the level of the biocontainment precautions required to isolate dangerous biological agents in an enclosed
The levels of containment range from the lowest biosafety level 1 to the highest at level 4, and Ministry of Health and Welfare and Korea Centers
for Disease Control and Prevention have specified these levels facility.
Biocontainment can be classified by the relative danger to the surrounding environment as biological
safety levels. As of 2006, there are four safety levels. These are called BSL1 through BSL4