Paris PROTOCOLS
From 2007.igem.org
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| - | '''Prepare to begin''' | + | '''Prepare to begin'''<br> |
<br> | <br> | ||
| + | ''This topic is adressed to all our informatics-physics-I'm-afraid-of-the-bench fellows. So, if you finally found the courage to dare the pipettes, PCRs and nicely smelling bacteria, welcome!'' | ||
<br> | <br> | ||
| - | + | * What a pipette is? [[(see image)]] | |
<br> | <br> | ||
| - | + | Pipettes dispense various volumes. The '''plunger button''' indicates the maximum volume (microliters) that the pipette is designed to handle. For example, P-20 will handle up to 20 microliters.<br> | |
| - | + | The '''digital volume indicator''' is read from top to bottom. For P-2, P-10, P-20, P-100, and P-200, black digits indicate microliters and red digits tenths and hundredths of microliters. For P-1000, red digits indicate milliliters and black digits microliters.<br> | |
<br> | <br> | ||
| - | + | What to do when you have it in you hand?<br> | |
| - | + | ||
<br> | <br> | ||
| - | + | -Hold the pipette in one hand (it doesn't bite...). With the other hand, turn the volume adjustment knob counterclockwise so the volume indicator is 1/3 revolution above the desired setting, then slowly turn clockwise until the indicator shows the desired volume.<br> | |
| + | -Attach a new disposable tip to the pipette shaft.<br> | ||
| + | -Press the plunger to the FIRST stop. This part of the stroke is the volume displayed by the indicator.<br> | ||
| + | -Holding the pipette vertically, immerse the tip a few millimeters into the sample.<br> | ||
| + | -Allow the pushbutton to return slowly to the UP position. Avoid to blurt out the plunger button abruptly : there are bulls appearing and your volume is false...<br> | ||
| + | -Ensure that the full volume of sample was properly drawn into the tip. <br> | ||
| + | -Withdraw the tip from the sample.<br> | ||
| + | -To dispense the liquid, gently touch the tip to the side of the receiving vessel, immersing the tip into liquid within the vessel. Press the plunger to the SECOND stop.<br> | ||
| + | -With the plunger fully pressed, withdraw the tip carefully, wiping residual drops against the vessel wall.<br> | ||
| + | -Allow plunger to return to the UP position.<br> | ||
| + | -Discard the tip by depressing the tip ejector button.<br> | ||
| + | <br> | ||
| + | Note down that different tips exist : ensure that you have the right one (labels will indicate you the size, etc.). It's better to use filter tips.<br> | ||
| + | To train, you can simply pipette water : it's important to know how much 1 µl is...<br> | ||
| + | <br> | ||
| + | ''To be continued...''<br> | ||
| + | <br> | ||
| + | <br> | ||
| + | * Growing bacteria in liquid medium <br> | ||
| + | <br> | ||
| + | -Light the Bunsen burner. It permits you to keep a 10 cm perimeter sterile et thus not to contaminate your future colonies. <br> | ||
| + | -Get a 50mL Falcon tube and put into 5 mL of LB medium. Add supplementary stuff if needed (antibiotics, metabolites, etc.).<br> | ||
| + | -Pick up a sterile toothpick. Use it to gather a single colony of cells (you know, a white point on your Petri dish...).<br> | ||
| + | -Place the toothpick with the colony into the solution.<br> | ||
| + | -Incubate overnight at 37°C with shaking (at about 200 rpm).<br> | ||
| + | <br> | ||
| + | Next morning, after a cup of coffee and a croissant, you can check up [[(see image 1)]].<br> | ||
Latest revision as of 23:19, 5 July 2007
Prepare to begin
This topic is adressed to all our informatics-physics-I'm-afraid-of-the-bench fellows. So, if you finally found the courage to dare the pipettes, PCRs and nicely smelling bacteria, welcome!
- What a pipette is? (see image)
Pipettes dispense various volumes. The plunger button indicates the maximum volume (microliters) that the pipette is designed to handle. For example, P-20 will handle up to 20 microliters.
The digital volume indicator is read from top to bottom. For P-2, P-10, P-20, P-100, and P-200, black digits indicate microliters and red digits tenths and hundredths of microliters. For P-1000, red digits indicate milliliters and black digits microliters.
What to do when you have it in you hand?
-Hold the pipette in one hand (it doesn't bite...). With the other hand, turn the volume adjustment knob counterclockwise so the volume indicator is 1/3 revolution above the desired setting, then slowly turn clockwise until the indicator shows the desired volume.
-Attach a new disposable tip to the pipette shaft.
-Press the plunger to the FIRST stop. This part of the stroke is the volume displayed by the indicator.
-Holding the pipette vertically, immerse the tip a few millimeters into the sample.
-Allow the pushbutton to return slowly to the UP position. Avoid to blurt out the plunger button abruptly : there are bulls appearing and your volume is false...
-Ensure that the full volume of sample was properly drawn into the tip.
-Withdraw the tip from the sample.
-To dispense the liquid, gently touch the tip to the side of the receiving vessel, immersing the tip into liquid within the vessel. Press the plunger to the SECOND stop.
-With the plunger fully pressed, withdraw the tip carefully, wiping residual drops against the vessel wall.
-Allow plunger to return to the UP position.
-Discard the tip by depressing the tip ejector button.
Note down that different tips exist : ensure that you have the right one (labels will indicate you the size, etc.). It's better to use filter tips.
To train, you can simply pipette water : it's important to know how much 1 µl is...
To be continued...
- Growing bacteria in liquid medium
-Light the Bunsen burner. It permits you to keep a 10 cm perimeter sterile et thus not to contaminate your future colonies.
-Get a 50mL Falcon tube and put into 5 mL of LB medium. Add supplementary stuff if needed (antibiotics, metabolites, etc.).
-Pick up a sterile toothpick. Use it to gather a single colony of cells (you know, a white point on your Petri dish...).
-Place the toothpick with the colony into the solution.
-Incubate overnight at 37°C with shaking (at about 200 rpm).
Next morning, after a cup of coffee and a croissant, you can check up (see image 1).
