LASA Biotech I - Midterm

Yes, there's redundant information on here. No, it's not by accident.

What is Biotechnology?

Using living organisms, or the products of living organisms, for human benefit to make a product of solve a problem

What are some examples of traditional biotechnology?

Fermentation, Selective breeding

What are some examples of modern biotechnology?

Genetic engineering to make insulin

What is genetic engineering?

The manipulation of an organisms genes by introducing, eliminating, or rearranging specific genes.

What are some career options in Biotechnology?

Sales, marketing, regulatory, legal, financial, human resources, administrative staff

What are some examples of biotech jobs that can be obtained with different degree type?

Postdoctorate - Scientist

Doctorate - Scientist

Master’s Degree - Research Associate

Bachelor’s Degree - Research Associate

Certificate - Biotechnician

High School Diploma - Lab Assistant

What is a MSDS/SDS?

Material Safety Data Sheet / Safety Data Sheet

Legally required technical document, provided by chemical suppliers, describing the specific properties of a chemical.

What are the 8 sections of an MSDS/SDS?

Chemical identity

Hazard ingredients/identity

Physical chemical charcateristics

Fire and explosion hazard data

Reactivity data

Health hazards

Precautions for safe handling and use

Control measures

What does the blue section of the NFPA diamond show?

Health Hazard

What does a 0 mean for health hazard?

Normal material

What does a 1 mean for health hazard?

Slightly hazardous

What does a 2 mean for health hazard?

Hazardous

What does a 3 mean for health hazard?

Extreme danger

What does a 4 mean for health hazard?

Deadly

What does the red section of the NFPA diamond show?

Flammability

What does the 0 mean for flammabiblity?

Will not burn

What does the 1 mean for flammability?

flash point > 200F

What does the 2 mean for flammability?

flash point >100F

What does the 3 mean for flammability?

flash point <100F

What does the 4 mean for flammability?

flash point <73F

What does the yellow section of the NFPA diamond show?

Reactivity

What does a 0 mean for reactivity?

stable

What does a 1 mean for reactivity?

unstable if heated

What does a 2 mean for reactivity?

Violent chemical change

What does a 3 mean for reactivity?

Shock and heat may detonate

What does a 4 mean for reactivity?

may detonate

What is PPE?

Personal Protective Equipment

What does the uncolored section of the NFPA diamond show?

Specific hazards

What does “OX” mean in the uncolored section of the NFPA diamond?

Oxidizer compound

What does “ACID” mean in the uncolored sectino of the NFPA diamon?

Acidic compound

What does “ALK” mean in the uncolored section of the NFPA diamond?

Basic compound

What does “CORR” mean in the uncolored section of the NFPA diamond?

Corrosive compound

What does “W” mean in the uncolored section of the NFPA diamond?

use NO WATER

What does LD50 mean?

Dose is lethal in 50% of test animals

What does LC50 mean?

Concentration in the air is lethal in 50% of test animals

Examples of PPE for handeling reactive substances?

Lab coats, goggles, gloves.

Tips for handling corrosive substances

Always slowly dilute strong acids or bases by adding water, store strong acids and bases apart from one another.

How should solutions be labeled in the lab?

Identity of the contents and its concentration, owners initials, date and time, owners class period.

What is a microcentrifuge used for?

to seperate samples based on desity by spinning at high speeds.

What is a micropipette used for?

used to measure and transfer liquids by drawing the liquid into the tip.

What is a seroligical pipette used for?

transferring militers of liquid from less than 1ml to 50ml.

What is a spectophotometer used for?

measure absorbance of light in the ultraviolet region of the specturm (~100-700nm). Require halogen light bulbs that emit ultraviolet light and require cuvettes that don’t absorb UV lights.

What is a gel electrophoresis apparatus used for?

to seperate nucleic acids and proteins based on their size and charge. Used to run and compare DNA samples.

What is a vortex used for?

mixes the contents by rotating the bottom of a tube rapidly

What is a thermal cycler (PCR) machine used for?

used for amplification of a specific section of DNA by PCR (polymerase chain reaction)

Where can a picofuge be found in the lab?

in a cabinet to the right of your group table

Where can a microcentrifuge be found in the lab?

in a cabinet to the right of your group table & in the small room on a table

Where can a micropipette be found in the lab?

Top left on group tables shelves

Where can micropipette tips be found in the lab?

bottom level of the shelves on the table

Where can microtubes be found in the lab?

Right bottom shelf on table

How do you use a micropipette properly?

Never lay/hold it upside down/sideways. Set to the desired amount using the black wheel located near the handle. To draw up liquid press the plunger down until the first stop, place the tip in the liquid, and slowly release the plunger. Releasing liquid: place the tip in the container, and press the plunger down until the second stop.

How do you read a P-10 micropipettor?

Limits are 1-10μl. Bottom value (under red line) reads the 1/10th number, middle value reads the 1s number, top value reads the 10’s number.

How do you read a P-100 micropipettor?

Limits are 1-100μl. Bottom value reads the 1s number, middle value reads the 10s number, top value (above red line) reads the 100s number.

How do you read a P-1000 micropipettor?

Limits are 1-1000μl. Bottom value reads the 10s number, middle value reads the 100s number, top value (above red line) reads the 1000s number.

If you were pipetting with a P-10 that was adjusted to 089 what amount would you be pipetting in μl and mL?

8.9μl, 0.0089ml

If you were pipetting with a P-100 that was adjusted to 089 what amount would you be pipetting in μl and mL?

89μl, 0.089ml

If you were pipetting with a P-1000 that was adjusted to 089 what amount would you be pipetting in μl and mL?

890μl, 0.89ml

Conversion factor from μl to ml

1ml = 1000μl

How do you balance a microcentrifuge?

Match the exact amount of liquid used in the microtube for a dummy microtube and place the two directly across from one another to balance themselves out.

What are the four ways cheese can be made? (names only)

Control/No treatment method

Starter Culture/Buttermilk Method

Rennin/Rennet Method

GE Chymosin Method.

Rennin/Rennet Method

Protease that breaks down the protein casein is retrieved from the cells lining the stomach of calves. Examples include asiago, brie, and most cheddars.

GE Chymosin Method

Rennin cheese-making gene is cut out of the cow’s stomach and inserted into fungus cells. Fungus cells then read the cow DNA and synthesize the rennin enzyme, called “chymosin”. Benefits: cheaper, faster, larger yields. Examples include Jack, mozzarella, most Swiss chesses, etc.

Control/No treatment method

Milk is left to age, exposed to aid and naturally occuring bacteria within the milk.

Starter Culture/Buttermilk Method

New batches of cheese are started with specific cultures of selected bacteria that contain the enzymes that curdle milk.

What bacteria is used in buttermilk as a starter?

Lactobacillus

Casein

Milk protein that denatures and falls out of the solution because of lactic acid. Denatured casein lumps are called curds.

Whey

Liquid remaining when curds are pressed together

Proteases

Cleade proteins, such as casein, into smaller fragments that will also fall out of the solution

How is lactic acid produced during cheese production?

The naturally occuring bacteria in milk uses lactose (milk sugar) as an energy source and produces lactic acid as a waste product.

Curd

Lumps of denatured casien that are pressed together to form cheese.

How to calibrate a pH meter

Rinse electrode with DI water and blot it with lint-free tissue, place the electrode into the frist buffer and press Cal to start the calibration and record the final vpH value. Repeat these steps for all your buffer solutions. To recall the calibration results, press and hold the Cal button.

How to calibrate an electronic balance

Press and hold Menu until the menu is displayed, release the button and the display [C.A.L] will appear - press yes to accept. [SPAN] will be shown - press yes to begin the span calibration. The specified mass blinks on the display. Press the specified span calibration mass on the pan or press No to select an alternate weight. [—C—] will blink while the reading is stored. The display will show [done] if the calibration was successful. The balance returns to the precious application mode and is ready for use.

Prokaryotic cells

Contains a cell wall made of peptidoglycan that protects and provides structure to the cell.

Cytosol: liquid within cells that contains biomolecules and chemicals used in the cells.

Plasma membrane made of phospholipid bilayer and proteins, regulates transport in and out of the cell.

Ribosomes: protein synthesis, RNA to Proteins.

RNA attached to the cell membrane/floating in the cytosol.

DNA free-floating in the cytosol.

Eukaryotic cells

Contains membrane bound organells.

Cytoplasm: cytosol + organelles, specific to eukaryotes.

Mitochondria: “powerhouse of the cell” because it converts energy stores in nutrients into ATP whic his used for energy within the call.

Lysosome: digest enzymes used for intracellular digestion.

Ribosomes: protein synthesis, made of RNA and protein.

Nucleus: contains DNA, genetic info.

Rough ER containts ribosomes, Smother ER doesn’t. Lipid synthesis and metabolism.

Golgi apparatus packages/modifies proteins for secretion from cells or for somewhere within the cell.

Function of the Nucleus

Contains the genetic information in eukaryotic cells and acts as the cell’s control center.

Function of the Rough ER

Ribosomes attached to its surface. Mostly known to be involved in the synthesis and production of proteins.

Extra: Secretes proteins made by bound ribosomes that are then moved to the transitional ER where they are wrapped in a transport vesicle to head to the Golgi apparatus.

Function of the Smooth ER

Mostly known to be involved in the production of enzymes and lipids.

Extra: Performs synthesis of lipids, metabolism of carbohydrates, detoxification of drugs and poisons, and stores calcium ions.

Function of the Golgi Apparatus

Packages proteins in little sacs called vesicles and distributes them around the cell to where they’re needed.

Extra: Modifies, stores, and sends proteins that come from the Rough ER.

2 sides to the Golgi Apparatus: cis and trans face. Vesicles enter via the cis face and depart via the trans face.

Also involved in the production of lysosomes.

Function of the plasma membrane

Made of a phospholipid bilayer where the head is hydrophilic and the tail is hydrophobic. Very flexible. Provides protection and a fixed environment inside the cell. Controls what enters and exits the cell.

Function of the Vacuole

Large vesicles that store many different things like food and water. Many unicellular eukaryotes have contractile vacuoles to pump water out of the cell.

Function of the mitochondria

Creates ATP for the cell to use via cellular respiration. Contains an outer membrane and inner membrane, making up its double membrane.

Extra: inner membrane consists of folds called cristar which is where most of the ATP production happens. The folds increase the surface area and thus the efficiency. Inside the inner membrane is the mitochondrial matrix which is the site of the Krebs Cycle.

Function of the chloroplast

Site of photosynthesis. Contains a double membrane and has a green pigment calles chlorophyll that allows for the absorbtion of photons.

Extra: Chloroplast is made of the stroma, or lliquid fillind of the chloroplast, and the thylakoids, flat sacs of membranes that allow for the absorption of light.

Function of the lysosome

Trash can of the cell.

Breaks down most foods, amino acids, and other molecules. Inside of lysosome is extremely acidic.

Extra: in charge of apoptosis, which is programmed cell death. When the lysosome bursts, the acid inside causes the cell’s death.

Differnence between animal and plant cells?

Plant cells contain cell walls made of cellulose that acts as a protective outer layer other than the membrane, plasma membranes, nucleus containing DNA, and Ribosomes. They do not contain Centrioles

Animal cells contain plasma membranes, nucleus containingDNA, centrioles, and ribosomes. They do not contain cell walls.

Nucleosome

DNA wrapped around histones

Extra: Helps to supercoil the DNA, resulting in a greatly compacted structure that allows for more efficient storage of genetic information.

Histones

Packages DNA in small packets that fit inside nucleosomes. Also serves as a mechanism to control gene expression.

Central Dogma

DNA —> RNA —> protein

DNA to RNA through transcription

RNA to protein through translation

Purpose of Replication

When a double-stranded DNA molecule is copied to produce two identical DNA molecules. The passing of genetics to the next generation.

Essential because whenever a cell divides, the 2 new daughter cells must contain the same genetic information as the parent cell.

Purpose of Translation

When information encoded in mRNA directs the addition of amino acids during protein synthesis.

3 components of a nucleotide

Deoxyribose Sugar, Phosphate, Nitrogen Base

Deoxyribose Sugar

5 carbon ring (pentose sugar). Each carbon is numberd clockwise starting from oxygen: 1’, 2’, 3’, 4’, 5’. Lacks OH on 2’ carbon.

Phosphate group

comes off the 5’ carbon

Nitrogenous base

attached to 1’ carbon: these vary for each of 4 types of nucleotides: adenine, thymine, cytosine, guanine (A, T, C, G)

DNA structure:

Double Stranded Helix

A - T: 2 H-bonds

C - G: 3 H-bonds

Complementary

Negatively charged sugar/phosphate backbone.

Antiparallel: 5’ to 3’, 3’ to 5’

Purines

One of the two nitrogenous bases.

Double rings, A and G.

Pyrimidines

One of the two nitrogenous bases.

Single rind, C and T (Uracil, U in RNA)

DNA base pairs

A - T, C - G

Adenine (A) to Thymine (T)

Cytosine (C) to Guanine (G)

RNA base pairs

A - U, C - G

Adenine (A) to Uracil (U)

Cytosine (C) to Guanine (G)

How are nucleotides linked together?

Covalently linked together in a chain through the sugars and phosphates

Purpose of DNA replication?

to produce 2 identical DNA molecules.