Term
| Where is the closest chemical shower located? |
|
Definition
| 2nd floor, G chem or O chem labs. |
|
|
Term
| If you were to get a chemical burn in your eye, how long would you rinse out your eye in the eye wash? |
|
Definition
|
|
Term
| What process must one go through when turning on an electrophoresis chamber for use? |
|
Definition
1. Place lid on chamber, make sure black matches black and red matches red
2. Plug in to outlet
3. Turn on Electrophoresis
4. Make sure bubbles are present |
|
|
Term
| What are the three main types of personal protective equipment one should wear when working with hazardous chemicals? |
|
Definition
1. Gloves
2. Goggles
3. Lab Coat |
|
|
Term
What are the 5 pipettes used in genetics Lab?
What volume can be measure out by each? |
|
Definition
p10- .05-10 microL
p20- 0.2-20 microL
p40- 05-40 microL
p100- 10-100 microL
p1000- 200-1000 microL |
|
|
Term
| If a person breaks or spills something, or is injured in lab, what is the first thing to do? |
|
Definition
|
|
Term
| What kind of toxin is polyacrylamide? |
|
Definition
|
|
Term
| Give 2 examples of clothing not to be worn in lab |
|
Definition
| Sandals, skirts, shorts, really long sleeves, dresses... |
|
|
Term
| What is the best way to prevent contamination of pipette tips? |
|
Definition
| Keep top closed as much as possible |
|
|
Term
| If you need to make a 2% agarose gel and you have .75 g agarose how much buffer is needed? |
|
Definition
|
|
Term
| What is the cutoff time for calling the TA if you need help? |
|
Definition
|
|
Term
| Give 3 examples of how DNA fingerprinting can be utilized in the real world |
|
Definition
1. Paternity
2. Crime Scenes
3. Genetic Disease |
|
|
Term
| Name some restriction enzymes |
|
Definition
| EcoRI PstI BamHI PvuII ScaI HindIII |
|
|
Term
| How are names given for restriction enzymes? |
|
Definition
1st letter of genus
1st 2 letters of species
Roman numerals indicating strain |
|
|
Term
| What is the average size of a recognition site for a restriction enzyme? |
|
Definition
|
|
Term
| What does RFLP stand for? |
|
Definition
| Restriction fragment length polymorphism |
|
|
Term
| Who discovered the structure of the DNA molecule? |
|
Definition
|
|
Term
| When detergent is added to the cells of onion or banana, what does it do to the cells? |
|
Definition
|
|
Term
| What are the 4 base pairs found in DNA, and which pair together? |
|
Definition
Guanine-Cytosine
Adenine-Thymine |
|
|
Term
| What is the backbone of DNA made from? |
|
Definition
|
|
Term
| What is the term used to identify breakage of the bonds found in the backbone of DNA? |
|
Definition
|
|
Term
| What is the site recognized by the EcoRI restriction enzyme? |
|
Definition
|
|
Term
| What is the chemical utilized in the gel to made DNA visible under UV light? |
|
Definition
|
|
Term
Common unit used for each of the following measurements:
Length
Mass
Amount
Concentration
Volume |
|
Definition
|
|
Term
| If you were asked to measure 300 microL of a substance, how many mL? |
|
Definition
|
|
Term
| If you utilized 30 mL of buffer, and 0.75 g agarose what percent gel are you making? |
|
Definition
|
|
Term
| What percent gel was used in the paternity lab? |
|
Definition
|
|
Term
| Name the person who first describe RFLP. |
|
Definition
| English geneticist, Alec Jeffries in 1985 |
|
|
Term
| How do restriction enzymes function in a lab? |
|
Definition
Restriction enzymes determine DNA characteristics.
They cut DNA into fragments which are then run in a gel to compare fragment length and discover the restriction sites.
They can be used in cases of murder, rape, incest... |
|
|
Term
| Bacteria from which EcoRi is derived? |
|
Definition
|
|
Term
| How are restriction enzymes used wiithin bacteria cells? |
|
Definition
They are bacterias most reliable defense mechanism.
They are used to cut out DNA and can be incorporated into the bacterias own DNA, which aids in things such as antibiotic resistance |
|
|
Term
| What is the maximum number of fragments that can be created from one restriction enzyme? |
|
Definition
| Depends how many times the sequence recognized by the enzyme appears |
|
|
Term
| What were the first 2 restrction enzymes utilized in the southern blotting lab? |
|
Definition
|
|
Term
| Why is it important to change pipette tips between samples? |
|
Definition
Contamination is very easily done
You change your tip so that you prevent contamination between samples. |
|
|
Term
| What does DNA fingerprinting tell you about DNA? |
|
Definition
| It tells where DNA has been cut and allows you to know what base pairs make up that individuals genetic character. |
|
|
Term
| What is the name of the process by which DNA segments are amplified? |
|
Definition
| Polymerase Chain Reaction PCR |
|
|
Term
| What is the distinguishing feature of restriction enzymes? |
|
Definition
| The ability to cut DNA and certain sites, and allow scientists to analyze the fragments. |
|
|
Term
| What are sticky ends of DNA? |
|
Definition
| When an enzyme cuts unevenly leaving some single strands hanging |
|
|
Term
| What are blunt ends of DNA? |
|
Definition
| When an enzyme cuts DNA evenly, so that there are no single strands present. |
|
|
Term
| How do you determine the number of times a restriction enzyme cleaves double stranded DNA? |
|
Definition
| It is determined by 4^n, n being the number of bp on the recognition site. |
|
|
Term
| What is the frequency for which EcoRI will cut DNA? |
|
Definition
|
|
Term
| What specific polymerase is typically used in PCR? |
|
Definition
|
|
Term
| What is the typical length of oligonucleotides? |
|
Definition
|
|
Term
| What tree components make up a gel? |
|
Definition
Agarose
1X TAE buffer
Ethidium bromide |
|
|
Term
| What is required for restriction enzyme activity? |
|
Definition
|
|
Term
| Why must one shake the gel tray during washing processes? |
|
Definition
| To keep the gel submerged |
|
|
Term
| List the order of items(from table up) in Southern blot to transfer DNA from gel to membrane. |
|
Definition
Plastic wrap
Gel
Membrane
Filter Paper
Paper Towels
Tray with large beaker |
|
|
Term
| How many restriction enzymes have been found to date? |
|
Definition
|
|
Term
| List the 6 major steps in Southern Blot following electrophoresis |
|
Definition
Denaturation
Blot transfer
membrane shielding
Detection
Color Development
Termination |
|
|
Term
| What chemical is required to be apart of the growth media for GFP to be expressed? |
|
Definition
|
|
Term
| Explain genetic transformation. What does it literally mean? |
|
Definition
Genetic transformation is the insertion of foreign DNA into an organism through a plasmid.
It literally means change caused by genes. |
|
|
Term
|
Definition
| Green Fluorescent Protein |
|
|
Term
| What is the genus and species of the jellyfish that GFP was isolated from? |
|
Definition
|
|
Term
| What antibiotic is the pGLO plasmid resistant to? |
|
Definition
|
|
Term
|
Definition
|
|
Term
| In Southern Blot analysis, why must you add the BCIP solution during the color development portion of the procedure? |
|
Definition
| So that the membrane will soak up that solution and when baked will allow your wells and bands to be visible. |
|
|
Term
| Why is southern blot a good tool for determining paternity? |
|
Definition
| It can be used to verify your results from RFLP electrophoresis. |
|
|
Term
| One the southern blot analysis is complete whats the best way to store the membrane? |
|
Definition
| Between filter paper in a dark place |
|
|
Term
| Why is it important to handle the filter paper with flathead forceps? |
|
Definition
| Because you can contaminate the sample if you touch it with your hands. Also, you can rip the membrane very easily if you use another tool |
|
|
Term
| When working with agar plates, why is it important to stack your plates upside down for incubation? |
|
Definition
| So that the condensation will not interfere with your results. |
|
|
Term
| If complete transformation has occurred in the pGLO experiment, what would cause the bacteria to glow under UV light? |
|
Definition
| The integration of the plasmid DNA containing the genes to make GFP and glow when turned on by arabinose into the bacteria |
|
|
Term
| In pGLO, what color will the bacteria glow under UV lights on arabinose plates if the experiment was successful? |
|
Definition
|
|
Term
| In pGLO, what color will the bacteria glow under UV lights on plates not containing arabinose if the experiment was successful? |
|
Definition
|
|
Term
| Which colonies of bacteria should the GFP gene be expresed in? |
|
Definition
| +pGLO grown on LB/amp/ara |
|
|
Term
| What is the purpose of having a control plate in your eperiment? For pGlO, what does this plate tell you? |
|
Definition
It is to compare your experimental results and to see if the bacteria had any natural ampicillin resistance.
The control plate told about the E coli before interacting with pGLO |
|
|
Term
| Why would it be possible for the +pGLO bacteria to grow on the plates containing ampicillin? |
|
Definition
| Because they received the ampicillin resistance gene from the plasmid |
|
|
Term
What is a plamid?
What is the average number of base pairs in a plasmid? |
|
Definition
A plasmid is an extrachromosomal piece of circular DNA that can replicate on its own.
Average number of bp=1,000-100,000 |
|
|
Term
| What is supercoiling and what causes it? |
|
Definition
| Form I DNA and when the DNA strands twist back on itself to form a very compact molecule, it is caused by the enzyme DNA gyrase |
|
|
Term
| What is the goal of the miniprep experiment? |
|
Definition
| To introduce students to the purification of plasmid DNA |
|
|
Term
| List the order of migration rates from fast to slow |
|
Definition
| Supercoiled>linear DNA>Nicked circles>dimers>trimers>tetramers |
|
|
Term
| T/F Plasmid DNA replicates separately of chromosomal DNA |
|
Definition
|
|
Term
| T/F Supercoiled plasmids exist only in the form of a dimer |
|
Definition
|
|
Term
| T/F DNA is a positively charged molecule |
|
Definition
|
|
Term
| T/F DNAse I is an endonuclease that creates nicks in plasmid DNA at specific sites? |
|
Definition
|
|
Term
| T/F In gel electrophoresis, supercoiled plasmids tend to migrate through a gel faster that other forms of plasmids |
|
Definition
|
|
Term
| How many base pairs are generally in a plasmid? |
|
Definition
|
|
Term
|
Definition
|
|
Term
|
Definition
|
|
Term
| What is a nick and what causes nicks in plasmid DNA? |
|
Definition
| A nick is a break in the phosphate bonds, and can be caused by chemicals, time, and DNAse I. |
|
|
Term
|
Definition
| Catenanes are multimers of plasmid DNA such as dimers, trimers etc |
|
|
Term
| What is SDS and NaOH used for in miniprep and how do they work? |
|
Definition
| They dissolve cell membranes and denature proteins. The high alkalinity aids in protein denaturation and causes the phosphate bonds of DNA to break. |
|
|
Term
| Why will degraded RNA migrate faster through an electrophoresis gel than DNA? |
|
Definition
| Because degraded RNA is smaller. |
|
|
Term
| Why is it important to remove RNA from the solutions during mini-prep and how is it removed? |
|
Definition
| RNA interferes with plasmid purification. RNAse removes it by degrading it. |
|
|
Term
|
Definition
| A vector is a plasmid that is used for propagation of genes |
|
|
Term
| What are vectors extremely useful for in molecular biology? |
|
Definition
| Vectors are useful for cloning. |
|
|
Term
| Is it possible for there to be more than one copy of plasmid DNA within a cell? |
|
Definition
| A cell can have multiple copies of plasmid DNA within it. |
|
|
Term
|
Definition
| Polymerase Chain Reaction |
|
|
Term
| What do you obtain when using PCR? |
|
Definition
| Many copies of a particular fragment of DNA from a tiny amount of starting material. |
|
|
Term
|
Definition
| Location for a specific allele |
|
|
Term
|
Definition
|
|
Term
| What are the three steps of PCR? |
|
Definition
Denaturation: melting of DNA, changing its shape
Annealing: Adding primers which make it ready to add nucleotides
Extension: Polymerase adds nucleotides |
|
|
Term
| From what bacteria is Taq isolated? |
|
Definition
|
|
Term
| Who invented PCR? what year? |
|
Definition
|
|
Term
|
Definition
| Exposed sheet of x-ray film |
|
|
Term
| Estimated number of genes with in human genome |
|
Definition
| ~30,000 according to handout |
|
|
Term
| What are the 4 deoxyribonucleotide triphosphates? |
|
Definition
|
|
Term
| What type of gel is used for sequencing reaction separation electrophoresis? |
|
Definition
|
|
Term
|
Definition
| The complementary nucleotides to the templates strand that experimentally is what you find out from reading/interpreting your gel. |
|
|
Term
|
Definition
|
|
Term
| What is the purpose of HLA? |
|
Definition
| Provides evidence about tissue compatibility |
|
|
Term
|
Definition
| Having 2 distinct cell lines within one person |
|
|
Term
| What biologicals can be used to test for HLA haplotypes? |
|
Definition
buccal mucosa
RBC
Thyroid
Hair |
|
|
Term
| What chromosome are the HLA genes ocated on? |
|
Definition
|
|
Term
| Why is it important to know a patient's HLA haplotype? |
|
Definition
To test for chimerism
To make sure they have compatible tissue type if they are going to receive a transplant of some sort. |
|
|
Term
| What is yeast's optimal temperature? |
|
Definition
|
|
Term
| What are the antigens on RBC's? |
|
Definition
|
|
Term
| What are the 6Rh antigens? |
|
Definition
|
|
Term
| Where are anti-Rh antibodies made? |
|
Definition
| They are made upon exposure |
|
|
Term
| What is the disease found in Rh+ newborns of a sensitized Rh- mother? |
|
Definition
|
|
Term
| What type of cell type of humans is used for karyotyping? |
|
Definition
|
|
Term
| What is needed for culturing human cells? |
|
Definition
buffer
CO2
Nutrients
Temperature ~37.5
PHA |
|
|
