Laboratory Manual in Bio117.: Cell & Molecular Biology For Marine Biology

Laboratory manual in bio117.
Cell & Molecular biology for marine biology 59

EXERCISE NO. 1
Introduction to Laboratory Tools
Answer Sheets
Name: Joana Jeanne L. Enteña Rating: __________

Laboratory Section: M45 Date: September 3, 2021
A. Write how many µl the following digital readout correspond to in each of the pipettes:
P20 P200 P1000
1 8. 0 1 4 3 8 8 0
18 µl 143µl 880 µl
0 7. 5 0 9 0 5 5 5
7µl 90 µl 555 µl
Write how the volume will be shown in the volume indicator of the following
micropipettes:
P20 P200 P1000
1
1 0 0 3 0 2 1 4
0
10 µl 130 µl 214 µl
8 5 0 7 8 0 3 8 9
8.5 µl 78 µl 389 µl
Laboratory manual in bio117. Cell & Molecular biology for marine biology
Guide Questions:
A. The Micropipette
1. Keeping the micropipette upright is a must when in use, why is this so?
- Avoids any liquid still present from running up into the inner workings (which can cause
corrosion).
- Avoids piston lubricant inside the pipette barrel from accumulating on one side only.
2. Give 3 advantages of using P1000 micropipette vs. 1ml glass pipette.

- A P1000 variable volume pipette for 100-1000µl adjusted with a Click - Stop digital
system for volume setting. Pipettes are color coded for easy identification.
- Micropipette is calibrated as per the standard of EN ISO 8655 for ensured accuracy &
precision andcalibration report is supplied.
- Glass pipettes are not highly accurate for volumes less than 1 milliliter (1 ml), but the
automatic pipettes are both accurate and precise
3. When adding liquid to an existing liquid, how is it done? Why?
- When transferring liquids with volumes greater than 5mL5mL, they can be poured
directly into vessels. Graduated cylinders and beakers have an indentation in their
mouth, so they can be poured controllably as long as the two pieces of glass touch one
another. If pouring from an Erlenmeyer flask, or transferring a liquid into a vessel
containing a narrow mouth, a funnel should be used. Funnels can be securely held with
a ring clamp, or held with one hand while pouring with the other. Precision instruments
that are designed to accurately and precisely transfer volumes in the microliter range.
B. The Autoclave
1. Give 5 items in the laboratory that should not be autoclaved.
1 Flammable, reactive, corrosive, toxic or radioactive materials
2 Household bleach
3 Any liquid in a sealed container.
4 Any material contained in such a manner that it touches the interior surfaces of the
autoclave.
5 Paraffin-embedded tissue.
2. Why is there a need to fill the reservoir with water?

- For this reason, various norms are now suggesting maximum impurity levels for steam
feed water of autoclaves and sterilizer used in the medical field. In the case of
benchtop autoclaves (e.g., dental autoclaves), care must be taken to clean the water
reservoir regularly, and refill it with freshly purified water.
3. Differentiate sterilization and decontamination.

- sterilization is (uncountable) the process of treating something to kill or inactivate
microorganisms while decontamination is decontamination. (uncountable) The process
of treating something to kill or inactivate microorganisms.
C. The Centrifuge Machine
1. Give 3 importance of using the centrifuge machine in cell biology research.

- One vital laboratory application of centrifuges is in blood separation. The separation of
blood into its component cells and proteins such as white blood cells (WBCs), red blood
cells (RBCs), and platelets can be achieved with the help of laboratory scale centrifuges.
- Another important laboratory application of the centrifuge is in the separation of DNA.
For this, DNA samples must first be purified and prepped for separation (usually through
the addition of buffers to it) and then placed in a centrifuge for a fixed amount of time.
- easy way to separate out substances, and has been shown to be better than membrane
filtration methods in certain situations where membrane filtration systems are used.
Centrifugation has been particularly beneficial in molecular biology research, where
ultracentrifuges are able to separate out very small molecules.
- Centrifuges are devices that can be employed to place objects in a rotational motion with
respect to a fixed axis. This results in the application of a force on the object which is
perpendicular to the axis of the rotation. In simple terms, the centrifuge is used to spin
an object in a circle in order to make it experience an outward force. The key
applications of the centrifuge are touched upon in this article.
2. How does temperature affect the efficiency of the centrifuge machine?
- During centrifugation, the temperature can be influenced by the rotor material’s thermal

conductivity. Rotors made of metal, such as steel and aluminum, have a high density
and high thermal conductivity. They transfer heat efficiently and get chilled quickly.
3. Why must 2 tubes of the same quantity or mass be placed opposite each other in the
centrifuge machine?
- If you are spinning more than two tubes, only the tubes directly opposite each other have

to be equal in mass. At high speeds, a centrifuge can easily become unbalanced
if equal masses aren’t located opposite each other in the rotor.
D. The Spectrophotometer
1. Give 3 importance of using the spectrophotometer in cell biology research.

- The spectrophotometer measures quantitatively the amount of light passing through a
compound in solution as a fraction of the light emitted by the machine
(“Spectrophotometry”)
- It is used to measure certain ingredients in a drug to make sure it is effective and safe
for consumers. You can measure bacterial growth, or diagnose a patient based on how
much uric acid is present in their urine.
- This instrument takes advantage of the regular light absorption and scattering patterns of
chemical structures (Lab Manual, p.19). Specifically, it detects compounds absorbing
light at selected wavelengths and produces a number corresponding to its absorption
(Alberts, 2004). Each compound absorbs and scatters this light more than others at
specific wavelengths.
2. Briefly discuss the principle involved in operation of the spectrophotometer.

- The spectrophotometer is an instrument which measures an amount of light that a
sample absorbs. The spectrophotometer works by passing a light beam through a
sample to measure the light intensity of a sample. These instruments are used in the
process of measuring colour and used for monitoring colour accuracy throughout
production.
3. Give 2 important steps on how to properly handle cuvettes.
- Avoid contact with clear sides of cuvettes with any hard surface (to avoid scratches).
- While filling the cuvettes, avoid spillage of the solution on the outer side of the cuvettes.

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Laboratory manual in bio117.

Cell & Molecular biology for marine biology 59

Name: Joana Jeanne L. Enteña Rating: __________

P20 P200 P1000

2. Give 3 advantages of using P1000 micropipette vs. 1ml glass pipette.

3. When adding liquid to an existing liquid, how is it done? Why?

1. Give 5 items in the laboratory that should not be autoclaved.

1 Flammable, reactive, corrosive, toxic or radioactive materials

3 Any liquid in a sealed container.

2. Why is there a need to fill the reservoir with water?

3. Differentiate sterilization and decontamination.

C. The Centrifuge Machine

1. Give 3 importance of using the centrifuge machine in cell biology research.

2. How does temperature affect the efficiency of the centrifuge machine?

- During centrifugation, the temperature can be influenced by the rotor material’s thermal

- If you are spinning more than two tubes, only the tubes directly opposite each other have

1. Give 3 importance of using the spectrophotometer in cell biology research.

2. Briefly discuss the principle involved in operation of the spectrophotometer.

3. Give 2 important steps on how to properly handle cuvettes.

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