biotech test
What does a restriction enzyme do, AND why do scientists use them?
A restriction enzyme is a protein that cuts DNA at specific sequences, known as recognition sites. Scientists use them to manipulate DNA for various purposes, such as cloning, gene editing, and DNA fingerprinting.
Look at the following DNA cuts—tell me what kind of ends" each of them have.
GCCAATTCGT CCCATTCG ATAGACTC
CGTTAAGCA GGGTATGC TATCTGAG
Sticky blunt sticky
Name two specific restriction enzymes and show the site where they seek along with
1. EcoRI - recognizes the sequence GAATTC.
2. HindIII - recognizes the sequence AAGCTT.
What do the letters in RFLP stand for?
Restriction Fragment Length Polymorphism.
What is an RFLP? How would you explain it to someone else?
variation in the length of DNA fragments produced by the digestion of DNA with restriction enzymes. A method used to compare DNA samples based on the pattern of fragments created when the DNA is cut.
Three uses for Restriction analysis technology (do repeat)
1. Genetic mapping.
2. Forensic analysis.
3. Paternity testing.
Describe a genetic map (shape, where you would find one and what information it provides) And how scientist use them
A genetic map is a visual representation of the arrangement of genes on a chromosome. It typically appears as a linear diagram and can be found in genetic studies or research articles. It provides information about the relative positions of genes and can help scientists understand genetic relationships and inheritance patterns.
Describe how to make an agar gel
To make an agar gel, you would mix agar powder with a buffer solution, heat it until it dissolves, then pour it into a mold to cool and solidify, creating a gel that can be used for electrophoresis.
What was the purpose of the PCR lab we completed in class?
The purpose of the PCR lab we completed in class was to amplify specific DNA sequences, making it easier to study and analyze them.
What unit of measurement do our micropipettors measure in, and what is that symbol?
Our micropipettors measure in microliters, which is represented by the symbol "µL."
Two ingredients of an electrophoresis gel?
1. Agarose.
2. Buffer solution.
What is the name of the final stain that we use in the DNA electrophoresis lab?
The final stain used in the DNA electrophoresis lab is called Ethidium Bromide.
Which fragments on a gel run will travel the farthest?
The fragments on a gel that are smaller in size will travel the farthest. Smaller fragments can move through the pores of the gel more easily than larger ones.
What is the charge of DNA, and which electrode will it travel to in an electrophoresis run?
DNA has a negative charge due to its phosphate backbone. During electrophoresis, DNA will travel toward the positive electrode (anode).
Explain the steps involved to final stain a gel ( include substance names, amounts, and time)
1. Prepare a staining solution of Ethidium Bromide at a concentration of about 0.5 µg/mL.
2. Immerse the gel in the staining solution for a day
3. After staining, rinse the gel with buffer to remove excess stain.
4. Visualize the gel under UV light.
What is the purpose of buffer
The purpose of the buffer in electrophoresis is to maintain a stable pH and provide ions that help conduct electricity, ensuring that the DNA fragments migrate properly through the gel.
What enzyme do we use to recombine DNA?
The enzyme used to recombine DNA is called DNA ligase. It joins DNA fragments together by forming phosphodiester bonds.
What does PCR stand for and what happens while in the thermal cycler (include temperature and stages)?
PCR stands for Polymerase Chain Reaction. While in the thermal cycler, the process involves three main stages:
1. Denaturation (94 degrees Celsius) where the DNA strands separate.
2. Annealing (60 degrees Celsius) where primers attach to the target DNA sequences.
3. Extension (72 degrees Celsius), where DNA polymerase extends the primers to synthesize new DNA strands.
If we increase the concentration of the agarose gel from.8% to 1%, what do we expect will happen to the migration of the RFLP's?
If we increase the concentration of the agarose gel from 0.8% to 1%, we would expect the migration of the RFLPs to slow down. Higher agarose concentrations create a denser gel, which hinders the movement of larger DNA fragments.
How to find parents using RFLP
To find parents using RFLP, scientists compare the DNA patterns of the offspring with those of the parents. By identifying shared fragments, they can determine biological relationships.
Purpose of instagene
The purpose of Instagene is to remove contaminants and inhibitors from the DNA sample, allowing for more accurate PCR results.
When completing your PCR lab, you had to centrifuge your material multiple times. What did this tell you about the mole size of DNA (small, medium, or larger) and explain why?
When completing your PCR lab and centrifuging your material multiple times, this suggests that the DNA is of medium to larger size. Centrifugation helps separate components based on size and density, indicating that larger DNA fragments might settle differently than smaller ones.
What materials are in the master mix/primer used in PCR labs? Each material of 5 and purpose
1. DNA polymerase - enzyme that synthesizes new DNA strands.
2. Primers - short sequences that provide a starting point for DNA synthesis.
3. dNTPs (deoxynucleotide triphosphates) - building blocks of DNA.
4. Buffer - maintains optimal conditions for the reaction.
5. MgCl2 (magnesium chloride) - cofactor that enhances the activity of DNA polymerase.