biotech test

What is the role of a restriction enzyme in DNA manipulation?

A restriction enzyme is a protein that cuts DNA at specific sequences, known as recognition sites.

Why do scientists use restriction enzymes?

Scientists use them to manipulate DNA for various purposes, such as cloning, gene editing, and DNA fingerprinting.

What type of ends do the following DNA cuts have: GCCAATTCGT, CCCATTCG ATAGACTC, CGTTAAGCA, GGGTATGC TATCTGAG?

Sticky, blunt, sticky.

What is the recognition sequence for EcoRI?

EcoRI recognizes the sequence GAATTC.

What is the recognition sequence for HindIII?

HindIII recognizes the sequence AAGCTT.

What do the letters in RFLP stand for?

Restriction Fragment Length Polymorphism.

What is an RFLP?

A variation in the length of DNA fragments produced by the digestion of DNA with restriction enzymes.

List three uses for restriction analysis technology.

1. Genetic mapping. 2. Forensic analysis. 3. Paternity testing.

Describe a genetic map.

A genetic map is a visual representation of the arrangement of genes on a chromosome, typically appearing as a linear diagram, providing information about the relative positions of genes.

What is the process to make an agar gel?

Mix agar powder with a buffer solution, heat until dissolved, pour into a mold to cool and solidify.

What was the purpose of the PCR lab completed in class?

The purpose was to amplify specific DNA sequences for easier study and analysis.

What is the unit of measurement for micropipettors?

Microliters, represented by the symbol 'µL'.

Name two ingredients of an electrophoresis gel.

1. Agarose. 2. Buffer solution.

What is the final stain used in the DNA electrophoresis lab?

Ethidium Bromide.

Which DNA fragments will travel the farthest in a gel during electrophoresis?

Smaller fragments will travel the farthest.

What is the charge of DNA and which electrode does it travel to in electrophoresis?

DNA has a negative charge and travels toward the positive electrode (anode).

Explain the steps involved in staining a gel with Ethidium Bromide.

1. Prepare the gel by placing it in a staining tray.

2. 124.75ml of water with .25ml of fast blast

3. Pour the solution over the gel, ensuring it is fully submerged.

4. Incubate for a day

What is the purpose of buffer in electrophoresis?

To maintain stable pH and provide ions for conducting electricity.

What enzyme is used to recombine DNA?

DNA ligase.

What does PCR stand for and what happens during the thermal cycler?

PCR stands for Polymerase Chain Reaction. It is a technique used to amplify specific DNA sequences. During the thermal cycler process, three main steps occur:

1. Denaturation: The double-stranded DNA is heated to aStages in the Thermocycler

1 - Denaturation 94C Separate the molecule

2 - Annealing 60C Primer attachment

3 - Extension 72C copies are made

What happens to the migration of RFLPs when agarose concentration is increased from 0.8% to 1%?

The migration slows down due to the denser gel hindering larger DNA fragments.

How do scientists use RFLP to find parental relationships?

By comparing DNA patterns of offspring with parents to identify shared fragments.

What is the purpose of Instagene?

To remove contaminants and inhibitors from DNA samples for accurate PCR results.

What does centrifuging DNA samples multiple times indicate about the DNA size?

It suggests the DNA is of medium to larger size, as centrifugation separates components by size and density.

List the materials in the master mix/primer used in PCR labs and their purposes.

Primer for the PV92 Gene - attache to start and end of the gene wanted

ATCG nucleotides - Needed to make new strands of DNA

Taq Polymerase - Needed to attach to primer and trigger the creation of the new DNA

Magnesium - required to activate the Taq polymerase

Dye - give color

Buffer - add volume