Lab 1 Questions and summary

Lab Safety in Genetics

1. Outline/state a FEW general rules of lab safety in the genetics lab.

Be Prepared: Always review your lab manual before class to understand the procedures
and objectives. This helps you stay organized and better equipped for lab activities.
Be Neat and Careful: Keep your work area clean and organized. Handle all equipment,
especially glass and sharp objects, with care. Clean up any spills immediately.
Wear Proper Clothing: Protect yourself by wearing appropriate clothing, such as lab
coats and closed-toed shoes. Avoid wearing anything that could be damaged by
chemicals.
Handle Chemicals Safely: Treat all chemicals as potentially dangerous. Wear protective
eyewear, and immediately wash off any chemicals that come in contact with your skin.
Know the Safety Equipment: Familiarize yourself with the location of safety equipment
like fire extinguishers, first aid kits, and emergency exits.
Follow Lab Etiquette: Never eat or drink in the lab and always seek permission before
using equipment or performing experiments. Always report any accidents or spills to
your instructor.

2. What are some of the main hazards that a student might encounter in a genetics lab?

In a genetics lab, students may encounter hazards such as open flames, UV light exposure, high
voltage from gel electrophoresis, chemicals like Fly Nap, hot objects, and corrosive microscopy stains.

Using Pipettors

3. Why are micropipettes so important in genetics?

Micropipettes are important in genetics because they allow for precise measurement
and transfer of very small volumes of liquids, which is essential for accurate
experimental results.

4. List the 4 main pipettor sizes and their range of volume. Which will we likely use the least?

1. P10: 0.5 to 10 µL
2. P20: 2 to 20 µL
3. P200: 20 to 200 µL
4. P1000: 200 to 1000 µL

The P10 is likely the least used since it is designed for very small volumes.
 5. Why do you use a disposable tip with the pipettors? What sizes of tips are available for
the pipettors?

Disposable tips are used with pipettors to prevent cross-contamination between

samples. Tips are available in various sizes, including 10XL, 20µL, 100µL, 200µL, and
1250µL.

6. Below is a display window for a P1000. The top number is red while the bottom two
numbers are black. What volume is the pipette set on?

1000µL

7. There are two stopping points on the pipettor plunger. How are these
stopping point used?

First stopping point: used to load tip with the correct volume of liquid.
Second stopping point: used to release the liquid from the tip

8. List some best practices when using the pipettor.

-Never point a pipette up. This may cause the liquid to run into the
pipette which could potentially break it
-Release the plunger slowly and carefully
-Use the proper size tip with the pipette
-Use a new tip for each different liquid
-Use the correct pipette for the volume that is to be dispensed

Centrifugation
9. What is centrifugation? Why do we use centrifugation in the genetics lab?

Centrifugation is a technique that uses centrifugal force to separate particles from a

solution based on their size, density, and other factors, and is used in genetics labs to
isolate cells or molecules efficiently.

10. Describe your understanding of centrifugal force and the things that influence it.

Centrifugal force is the outward force exerted on particles in a rotating system, pushing
them away from the center of rotation. Its strength depends on the rotation speed
(higher speeds increase the force), the distance of the particles from the center (greater
distances result in stronger forces), and the densities of the particles (greater density
differences enhance separation).
 11. What size of tubes are available for common use in the genetics lab with a table top
microcentrifuge?

1.5 mL tubes
2.0 mL tubes
0.5 mL tubes

12. When you load the centrifuge with the appropriate tube, what direction should the
hinge be pointing?

The hinge should be pointed upward.

13. What are some important things that you should insure when working with a
centrifuge?

When working with a centrifuge, it is important to ensure that tubes are balanced
symmetrically to prevent imbalances, securely closed to avoid spills, and that the rotor is
properly installed. Additionally, the centrifuge should be clean and free of debris, with
correct speed and time settings for your protocol.

Gel Electrophoresis

14. Why do we do agarose gel electrophoresis? What is agarose? How is it different than
plain agar (use internet)?

Agarose gel electrophoresis is used to separate and analyze DNA, RNA, or proteins based on
their size and charge by passing them through a gel matrix. Agarose, a purified
polysaccharide derived from seaweed, forms a gel with defined properties suited for this
purpose. It differs from plain agar, which contains both agarose and agaropectin, making it
less refined and more suited for microbiological media rather than electrophoresis.

15. How do we generally visualize DNA in an agarose gel? Is there a dye? Is there a
special light? Describe these features.

To visualize DNA in an agarose gel, we use DNA-binding dyes such as ethidium

bromide or SYBR Green. Ethidium bromide fluoresces under ultraviolet (UV) light,
showing DNA bands as bright orange against a dark background. SYBR Green, fluoresces
under blue light, producing green bands visible on gel imaging systems.

16. What size of particle runs faster in an agarose gel? Large or small molecules of DNA?
 In an agarose gel, smaller molecules of DNA run faster than larger molecules because they can
navigate through the gel matrix more easily.

17. What is a DNA ladder (use internet) and why is it included in a gel?

A DNA ladder, also known as a DNA marker, is a mixture of DNA fragments of known
sizes used as a reference in gel electrophoresis. It is included in a gel to help estimate the
size of the DNA fragments in the sample by comparing their distance traveled to the
bands in the ladder.

DNA quantification
18. What is the optimum absorption wavelength (in nm) for double stranded DNA?

260nm

19. Why does DNA fluoresce under UV light?

DNA fluoresces under UV light because it contains nitrogenous bases (adenine, thymine,
cytosine, and guanine) that absorb UV light and re-emit it as visible fluorescence. This
property is utilized with fluorescent dyes, which bind to DNA and enhance its visibility
under UV light.

20. At 260nm, how much DNA (ng/ul) is 1 unit of absorbance equivalent to?

At 260 nm, 1 unit of absorbance is equivalent to approximately 50 nanograms per

microliter (ng/µL) of double-stranded DNA.

21. Based on the equation: [conc. of DNA] = Absorbance (260nm) * 50 ng/ul * dilution factor.
What is the concentration of dsDNA given the absorbance value of 0.70 at 260nm and a
dilution factor of 10?

Absorbance = 0.70
Dilution Factor = 10

0.70 x 50 ng/µL x 10 = 350

The concentration of dsDNA is 350 ng/µL.

Insert a photo of the filter paper color “wheel” that you made from pipetting.

Lab Summary (Provide a summary of this lab that addresses the main points of the lab. Use
complete sentences)

In Lab 1, I focused on essential lab techniques and protocols, starting with an introduction to
proper lab safety. This included understanding the importance of wearing appropriate lab attire,
safely handling chemicals, and correctly handling lab equipment. I gained hands-on experience
by practicing pipetting, which refreshed my memory from last semester on how to properly
measure and transfer liquids in a safe and effective way. Additionally, we went over what
agarose gel is, and the purpose it serves in the process of gel electrophoresis when analyzing
different samples of DNA. In this discussion, we also talked about how and why we use DNA
ladders in gel electrophoresis which was a nice refresher from what I had previously learned in
BIO 1020. Finally, we ended the lab by learning about spectrophotometry, a method for
measuring the absorbance of light by a sample, which is crucial for quantifying concentrations
of various substances. This technique involves using a spectrophotometer to detect how much
light is absorbed at specific wavelengths, providing valuable data for analyzing samples. This
was a technique I had not previously heard of in any of my science classes so it was definitely
interesting to learn all of the components involved in the process. All in all, this lab reinforced
my understanding of laboratory safety protocols and the use of common laboratory equipment,
preparing me well for future lab work.