1How do you find the dilution ratio?
volume moved/final volume
2How do you find the dilution between two different solutions?
Solve for the dilution ratio, then put it to the power of x - x being how many times it was diluted from the stock solution
3What is the equation for finding the concentration of a solution?
C1V1 = C2V2
4What is C1?
Concentration of stock
5What is V1?
Volume of the stock you need to add to the diluent
6What is C2?
The final/target concentration
7What is V2?
The final volume (stock added + diluent)
8How do you go from a standard unit to the mili-prefix?
Divide by 1000
9How do you go from a mili-prefix to the standard unit?
Multiply by 1000
10How do you go from a standard unit to the micro-prefix?
Divide by 10^6
11How do you go from a micro-prefix to the standard unit?
Multiply by 10^6
12What is a standard curve?
A curve that shows the relationship between two quantities
13What is the R^2 value?
Measures the error between the data and line - the closer to 1 the more accurate the data
14What is a Bradford Assay?
A colorimetric method used to measure the concentration of protein in a sample by observing the color change of a dye
15What color will the solution be when there is no protein concentration in a Bradford Assay?
Brown
16What color will the solution be when there is protein concentration in a Bradford Assay?
Blue - continues to get darker as protein concentration increases
17How does the Bradford Assay work?
The dye is bound to the protein so it will only show up when there is protein present
18What happens when there is too much substrate in a Bradford Assay?
It will plateau because it is supersaturated - no more change or sign of change
19What is an enzyme?
Enzymes are proteins that act as biological catalysts
20What do enzymes do?
Rather than being consumed, they help speed up the reactions
21How does an enzyme work?
A specific substrate attaches to an active site on the enzyme, and once bound, an enzyme-substrate complex is formed where the activation energy of the reaction is lowered and promotes rapid progression, then a product is released
22What is a hallmark property of enzymes?
They remain unchanged by the reactions they catalyze
23What is the lock and key idea?
An enzyme is a lock, and it will only work for a certain shaped key (substrate)
24What was the enzyme we used in our lab?
Beta-Galactosidase (B-Gal)
25What was the substrate we used in our lab?
ONPG
26Why did we need E. coli in our lab?
We needed it to produce the enzyme (B-Gal) - we lysed it to disrupt the cell membrane and release intracellular contents (B-Gal)
27What does B-Gal do?
Catalyzes hydrolysis of lactose into galactose and glucose
28How do you calculate the activity of something?
Absorbance/elapsed time (in seconds) x 100
29What does DNA contain?
Instructions for the development of an organism
30What is the central dogma?
The concept that genetic information flows only in one direction, from DNA, to RNA, to protein, or RNA directly to protein
31What is DNA composed of?
A double helix structure and nucleotides
32What are the four building blocks of DNA (nulceotides)?
Adenine, Thymine, Cytosine, and Guanine (Uracil found in RNA and takes place of Thymine)
33What is special about the two strands in DNA?
They are complimentary (A pairs with T, and C pairs with G) and they are antiparallel, twisting around itself
34What is coding DNA?
The type of DNA in the genome that encodes for protein-coding genes
35How much of the human genome is made up of coding DNA?
1%
36What is noncoding DNA?
DNA that does not encode for protein-coding genes
37How much of the human genome is made up of noncodign DNA?
99%
38What are STRs?
Short tandem repeats where a DNA sequence repeats itself many times (always 4 nucleotides long)
39Where are STRs found in?
The noncoding region of the genome that determines physical traits- will be on everyone's chromosomes in the same spot with differing amounts of repeats
40What is a Polymerase Chain Reaction (PCR)?
A method used for making billions of copies of a specific DNA sequence
41Why do researches use PCR?
So they can easily build up large quantities of genetic material for experimental use
42What is in the master mix of PCR?
DNA template, primers, DNA polymerase, Deoxyribose (dNTPs), reaction buffer, and PCR grade water
43What occurs during Cycle 1 of PCR?
Template denaturing, primer annealing, then elongation period
44What occurs during template denaturing in PCR?
The denaturtion of DNA results in the hydrogen bonds between complementary base pairs being broken yielding only single stranded DNAs
45What occurs during primer annealing in PCR?
The annealing of short pieces of DNA called primer. Specific to the sense or antisense strands bind via hydrogen bonds
46What occurs during the elongation period of PCR?
Elongation using polymerase, begins by adding dree dNTPs to the ends of the primer one at a time in the 5' to 3' direction to mske a double stranded DNA
47What occurs during Cycle 2 of PCR?
Primers will bind to sigle stranded DNA formed via previous extension and the amplicon (short fragment you are trying to amplify) will form when the polymerase extends fro the forward primer on a strand that was generated by amplification from the reverse primer or vice versa
48Why do we run the gel in electrophoresis from negative to positive?
DNA carries a negative charge (due to the phosphate backbone) so it is attracted to the positive end
49What happens to the smaller fragments in electrophoresis?
They will travel faster and go further
50What happens when the gel is denser in electrophoresis?
All the particles, regardless of size, will move slower
51What is heterozygous?
Two different versions of the gene inherited from parents
52What is homozygous?
Two identical alleles of the gene inherited from parents
53How will homozygous genes show up in gel?
They will show up as one band
54How will heterozygous genes show up in gel?
They will show up as two bands
55What do we call two lineages stemming from the same branch point?
Sister taxa