How to Pipette

Using a pipette is an essential part of working in a science laboratory. Pipettes,

specifically micro-pipettes, are used for accurately measuring and transferring
small amounts of solutions from on container to another. This is useful when
creating a dilution series, plating cell samples. etc.

Drawing up Volumes of Liquid with Pipettes

1. Use pipettes to accurately measure small volumes of liquid. Pipettes function
as tiny straws that suck up a liquid into the vacuum of its internal holding space. If
you wish to extract a very precise, yet small, volume of liquid, a pipette is the tool
you’ll likely use.

2. Keep the pipette tip away from the bottom of the vessel. When you want to
draw up a volume of liquid into the pipette, make sure you keep the tip of the
pipette elevated away from the bottom. Otherwise, you might cause a seal to occur,
which can create air bubbles.

3. Draw up liquid with the reverse pipetting technique. This method is only

possible with air displacement pipettes. They draw up the liquid and disperse with
any air bubbles or other forms of aspiration and is especially effective for drawing
up and dispersing foamy or viscous liquid samples.

4. Release liquid with the reverse pipetting technique. Once you’ve drawn up

the liquid, you’ll need to depress the plunger to the first stop to release the liquid
out of the pipette. Then you can discard the remaining liquid by depressing the
plunger button to the second stop.

Using Pipettes to Disperse Volumes of Liquids

1. Disperse volumes of liquid with pipettes. Pipettes are often used in labs—such
as chemistry, biology, or medical labs—to transport precise quantities of liquids or
other soluble materials. There are various types of pipettes, both manual and
mechanical, that can measure out a volume of some liquid and transport it to
another location for distribution.
2. Measure volumes accurately. Since pipettes are intended for use with smaller
volumes, they are typically very accurate when it comes to measuring these small
amounts of liquid. Both manual and mechanical pipettes are known for their
accuracy when it comes to measurements.

3. Use the forward pipetting technique. This technique is most commonly used

to expel an exact volume of liquid from the pipette. Mastering the technique means
that you will be able to repeat the procedure with exactly replicated results each
time.

4. Disperse the liquid with the forward pipetting technique. Depress the pipette
plunger to the first stop so that liquid is drawn up into the receiving vessel. Then
you’ll need to depress the pipette button a second time to the second full stop in
order to expel the desired amount of liquid from the pipette tip.

Cleaning Pipettes
1. Clean the outside with mild soap. Rinse off the exterior of the pipette with
water and apply some mild soap or detergent. Rinse the soap off with running
water, then use a clean cloth or paper towel to wipe off any remaining residue.

2. Clean the interior with distilled water and alcohol. If possible (depending on
the type of pipette being used), disassemble the pipette and clean the interior. Rinse
all the disassembled parts in distilled water, then soak each part in 10% isopropyl
alcohol. This will help remove any contaminants that may be contained inside.

3. Use a strong detergent to rinse off radioactive substances. For any

radioactive substances, you should rinse the pipette with a strong detergent like
Deconex. After, try rinsing the pipette again several times with distilled water.

4. Boil pipette pieces and rinse them with water to remove nucleic acids. For
nucleic acids, you should boil the pipette pieces in glycine/HCl buffer for ten
minutes. Then rinse the pipette pieces with distilled water, rather than using
alcohol to clean them.
5. Use a strong detergent solution to rinse off protein particles. For proteins,
you should rinse the contaminated pieces with a strong detergent solution, not
alcohol. Then rinse the pipette pieces with distilled water.

6. Allow to fully air dry before reusing. Once you have finished cleaning your
pipette, you should let it completely air dry before reassembling and using it again.
If you don’t, your volume measurements could be off, even if just minutely, from
the leftover liquid inside.

How to prepepare a chemical solution

Fill the volumetric flask about halfway with distilled water or deionized water
(aqueous solutions) or other solvent. Transfer the solid to the volumetric flask.
Rinse the weighing dish with the water to make certain all of the solute is
tranferred into the flask. Stir the solution until the solute is dissolved.

How to use a burette

1. Rinse the burette with the standard solution to be used, and align burette tube
vertically.
2. Fill the burette slightly above the zero mark. To prime the stopcock, drain the
burette no further than the nominal capacity.
3. Refill the burette with titrant free of air bubbles to approx. 5 mm above the zero
mark.
 4. Drain liquid to set the zero point accurately. Important: Meniscus must be read
at eye level (parallax-free level). Automatic burettes: Fill to approximately 5 mm
above the zero mark This is adjusted automatically after air release.
5. Wipe off any drops adhering to the discharge tip.
6. Open the stopcock and slowly add titrant to the sample (containing the
indicator). The discharge tip must not touch the wall of the vessel. Keep swirling
the sample vessel lightly while adding titrant, or place it on a magnetic stirrer.
7. Read the discharged volume at eye level.
8. Any drops remaining on the tip of the stopcock should be wiped against the
vessel wall and rinsed down. It is part of the titrated volume.

How to use an autoclave

Packaging and Loading

 Only designated individuals should be allowed to set and/or change

parameters for the autoclaves.
 Before using the autoclave, check inside for any items left by the previous
user that could pose a hazard.
 Clean the drain strainer before loading the autoclave.
 Always place items in a secondary container.
 Do not overload or package bags too tightly. Leave sufficient room for
steam circulation. If necessary, place container on its side to maximize steam
penetration and avoid entrapment of air.
 Use only autoclavable bags to package waste.
 Do not allow bags to touch the interior walls of the autoclave to avoid
melting of plastic.
 Ensure sufficient liquid is packed with contents of autoclave bags if dry.
 Place soiled glassware and lab ware in secondary containers and autoclave
them in the solids cycle. Do not fill containers more than 2/3 full with liquids.
Loosen caps or use vented closures.
 In case of clean glassware and wrapped instruments, lay them in a secondary
container before autoclaving in wrapped goods cycle.
 For secondary containment, use autoclave trays made out of polypropylene,
polycarbonate or stainless steel. The trays should have a solid bottom and sides to
contain the contents and catch spills.
 Choose appropriate cycle for the material.  Incorrect selection of cycle may
damage the autoclave, cause liquid to boil over or bottles to break.
 Start your cycle and fill out the autoclave user log.  A completed cycle
usually takes between 1 to 1.5 hours.
 Check chamber/jacket pressure gauge for minimum pressure of 20 pounds
per square inch (psi).
 Close and lock door.
 Check temperature for 250⁰F (121⁰C) every load.
 Do not attempt to open the door while autoclave is operating.

Unloading

 Ensure cycle has completed and both temperature and pressure have
returned to a safe range.
 Wear PPE described above, plus an apron and face shield if removing
liquids.  Stand back from the door as a precaution and carefully open door no more
than 1 inch. This will release residual steam and allow pressure within liquids and
containers to normalize.
 Allow the autoclaved load to stand for 10 minutes in the chamber. This will
allow steam to clear and trapped air to escape from hot liquids, reducing risk to
operator.
 Do not agitate containers of super-heated liquids or remove caps before
unloading.
 Place liquids in an area which clearly indicates the items are “hot” until the
items cool to room temp.
 Allow autoclaved materials to cool to room temperature before transporting.
Never transport superheated materials.
 Place cooled autoclaved biohazard bag into regulated medical waste box. 
Autoclaved infectious liquids may be disposed of into the sanitary sewer.

How to Use a Centrifuge

Security
Place the centrifuge in a secure place. It should not be in any danger of being
knocked off a table, or pulled off by a person tripping over a loose cord. The
centrifuge also has to be on a flat, sturdy surface so the vibration it creates when
it is running is kept to a minimum. If the machine wobbles excessively, turn it off
immediately in case it is malfunctioning or badly loaded.

Loading

Balance the load. If you have only one sample, for example, load another tube on
the other side directly opposite the sample that has an equivalent load in it. It is
important to balance it by mass, instead of simply by volume, as is recommended
by Stanford University's centrifuge usage tips. If the sample is more dense than
water, for example, you have to compensate by adding more density or volume to
the balancing tube.

Opening and Closing

Ensure that the lid is properly closed when you finish loading the centrifuge. In
addition, never open a centrifuge when it is operating, because even though the
machine may turn off, the residual energy can continue to spin the samples at a
high speed and the samples, or even the rotor itself, if it is broken, can fly off at
dangerous speeds.

How to Use an Electronic Balance

Electronic balances have become standard equipment for many high school and
college chemistry departments. They allow the user to quickly and accurately
measure the mass of a substance to a level of accuracy impossible for traditional
balances to achieve. This is especially important in experiments that require
precise amounts of each substance to achieve the desired results. The popularity
of the electronic balance is also due to its extreme ease of use for any skill level.

Place the electronic balance on a flat, stable surface indoors. The precision of the
balance relies on minute factors and wind, shaky surfaces, or similar forces will
cause the readings to be inaccurate.
Press the "ON" button and wait for the balance to show zeroes on the digital
screen.

Use tongs or gloves to place the empty container you will use for the substance to
be measured on the balance platform. Fingerprints and other greases from your
hands add mass and must be avoided for accurate measurements.

Press the "Tare" or "Zero" button to automatically deduct the weight of the
container from future calculations. The digital display will show zero again,
indicating that the container's mass is stored in the balance's memory.

Carefully add the substance to the container. Ideally this is done with the
container still on the platform, but it may be removed if necessary. Avoid placing
the container on surfaces that may have substances which will add mass to the
container such as powders or grease.

Place the container with the substance back on the balance platform if necessary
and record the mass as indicated by the digital display.