Bio 121 Lab: Exam 2 Guide
A dihybrid cross consists of 2 heterozygotes in the P generation. What i expected phenotypic ratio of their offspring?
*dihybrid* = 4 numbers = 9:3:3:1 9= Dominant/Dominant 3= Dominant/Recessive 3= Recessive/Dominant 1= Recessive/Recessive
What will be be the phenotypic ration of offspring in a monohybrid cross of 1 parent that is Heterozygous and 1 parent that is homozygous recessive?
*monohybrid= 2 numbers* Aa x aa = 1:1
can grow in the presence of the antibiotic ampicillin
+pGLO
Given the following- list all the condition sets where you would expect to see colony Growth.
-pGLO LB +pGLO LB/amp +pGLO LB/amp/ara * no growth in -pGLO LB/amp*
On the test, we will be told a marker was found. A reaction happened. Means?
-reaction means a marker was found. A blood means A marker B blood means B marker. AB is A and B marker. Antisera identifies the marker. Antisera A finds the A marker etc. 4 reactions: A, B, A+B, none=O (recessive)
What is the condition we observed in blood type where 2 dominant alleles can be expressed at once?
1. Codominance AB blood type or Genotype I^A I^B - means we have on our red blood cells, we have the A shaped marker and the B shaped marker
What blood type is expressed as dominant? Recessive? Codominant? (make sure to note which answer is which type of expression)
1. Dominant blood type is either A or B blood 2. Recessive blood type is O blood 3. Codominant blood type is AB
Define electrophoresis and how does it manipulate DNA in a way that is useful to us?
1. Is the technique used to separates DNA fragments based on size 2. this technique is useful to us to because it can identify individuals or can identify changes in DNA sequences (such as mutations)
Name the 3 genes contained on the pGLO plasmid
1. bla, araC, GFP
Name an describe the function of ONE of the genes contained in the pGLO plasmid.
1. bla- produces the enzyme for the bacteria to grow in the presence of ampicillin 2. araC- the promoter gene that is turned on in the presence of the sugar arabinose in the environment 3. GFP- produces the fluorescent pigment
Describe 1 similarity between the 2 types of bacteria used in this lab. -pGLO bacteria (untransformed) and +pGLO (transformed)
1. both are HB101 non-virulent strains of E. coli bacteria 2. both need LB agar to survive 3. both replicate at 37°C and take overnight to replicate enough for us to visualize colonies
What procedures do the cells require to undergo transformation
1. competence 2. stressing out the cells 3. letting them recover
What is a competent cell? And what did we do to make our cells competent?
1. competent- the cell membrane has holes in it that allow the plasmid to enter the cell. 2. to make them competent, we added calcium chloride, we heat shocked them and ice bathed them. page 131
Describe 2 difference between the 2 types of bacteria used in this lab. +pGLO bacteria (transformed)
1. contains plasmid 2. can survive in the presence of ampicillin and glows if there is arabinose in the agar
Describe 2 difference between the 2 types of bacteria used in this lab. -pGLO bacteria (untransformed)
1. does not contain plasmid 2. is killed by ampicillin
What is necessary for the cell to take up plasmid?
1. gave CaCl2 which poked holes in the cell membrane 2. heat shocking which stresses out the bacterial cell which so they will suck up the juice from the environment which ours had the plasmid.
There were a total of 400 corn kernel counted (exactly) by Jack and Jill in the lab last week. 311 of the kernels were purple and 89 were yellow. What is the EXPECTED phenotypic ratio of the offspring? (chai square test)
1. only 2 colors so its monohybrid = 2 numbers 2. Either 1:1 or 3:1 3. Based on the question, its 3:1
There are multiple conditions necessary for the E. coli to GLOW. Names at least ONE unique condition that would enable the bacteria to GLOW.
1. pGLO plasmid 2. UV light 3. Presence of the sugar
Define the concept of DNA fingerprinting. What is its purpose? What are the 4 samples that we observed in our experiment?
1. purpose is to identify DNA samples from the same individual. 2. sample from crime scene and 3 suspects.
There are multiple conditions necessary for the E. coli to Glow. Name at least ONE unique condition that would enable the bacteria to GLOW.
1. the presence of the sugar in the environment 2. the UV light
Which of the genotypes above indicates resistance to malaria? List all that apply 1. Homozygous Dominant 2. Heterozygous Recessive 3. Homozygous Recessive
2. Heterozygous Recessive 3. Homozygous Recessive - the environment keeps it around
Name the genotype that has sickle cell anemia. 1 . HbA/HbA 2. HbA/HbS 3. HbS/HbS
3. HbS/HbS also aa or q2
Ideal temperature for the growth of E.coli?
37 degrees C 98.7 degrees F
Know how to construct a monohybrid punnet square
4 square box with either male allele or female allele on the top or side cross multiply
genetic drift
A change in the allele frequency of a population as a result of chance events rather than natural selection.
natural selection
A natural process resulting in the evolution of organisms best adapted to the environment.
What is a plasmid and what are general characteristics of plasmids?
A small circular self-replicating DNA molecule that is separate from the bacterial chromosome
What will be the phenotypic ratio of offspring in a monohybrid cross of one parent that is Heterozygous and one parent that is homozygous recessive?
Aa x aa = ratio is 1:1
gene pool
All the genes, including all the different alleles for each gene, that are present in a population at any one time
Prevents growth of untransformed bacteria on LB/amp plates
Ampicillin
Genotypes vs phenotypes
An organism's genotype is the set of genes that it carries/ genetic makeup. An organism's phenotype is all of its observable characteristics/ physically expressed traits Blood- Geno is I^A; Pheno is A or B
Sugar (ose) that turns on GFP gene ( enables the +pGLO bacteria to glow)
Arabinose
derived character in only one taxon
Autapomorphy
Predictions of phenotypic ratios of offspring results of a monohybrid or a dihybrid cross, and given ratios of offspring, predict parental genotypes
Basically, the information in a question will be given and we need to set up a punnet square to answer the pheno and geno types -Heterozygous x Heterozygous = Mono-(AaxAa) is 3:1; Dihyb-(AaBbxAaBb) is 9:3:3:1. Heterozygous x Homozygous= Mono-(Azxaa) is 1:1; Dihyb- (AaBbxaabb) is 1:1:1:1
Compare and contrast population bottle necks and the founder effect. These are examples of _____?
Bottle necks: catastrophic event/ kills a huge portion of the population Founder effect: when individuals are separated from the original population and start a new population Both are examples of genetic drift due to chance
pokes holes in the cell membrane so plasmid can enter the cell
CaCL2 or calcium chloride
evolution
Change in a kind of organism over time; process by which modern organisms have descended from ancient organisms.
Define what it means that a bacterial cell is competent
Competence means the cell membrane has holes to allow the plasmid to enter the cell- holes are created by CaCI2
Are the colored pigments on a gel that are seen with our naked eye that show up after performing electrophoreses bands of DNA
DNA bands either have to be stained or some other treatment to be seen after electrophoresis. the pigments seen on the gel directly after electrophoresis are just the loading dye.
What is the charge of DNA - therefore, DNA moves toward which charge
DNA has a negative charge. DNA then moves towards positive charge.
Cladogram
Diagram that shows the evolutionary relationships among a group of organisms
species of bacteria used in the experiment
E. coli
Device used to generate a difference in electrical charge
Electrophoresis chamber
In regard to blood type, a person with type AB blood can donate to both type A and type B individuals. T/F
False If someone receives a blood type, they can't receive something different than they already have. If a person with A blood received AB blood, the body would recognize the B as foreign and attack it and vice versus.
Larger DNA fragments travel faster than smaller ones? T/F
False, smaller DNA would land further from their starting point.
gene that produces a pigment that will fluoresce under UV light
GFP
What strain of E.coli was used in the experiment and why
HB101 it is a non-virulent strain
Which of the genotypes indicates resistance to malaria and also sickle cell anemia symptoms? 1. Homozygous Dominant 2. Heterozygous Recessive 3. Homozygous Recessive
Heterozygous Recessive (sickle cell anemia is recessive disorder) sickle cell is genetic and malaria is environment
Heterozygous x heterozygous Mono (Aa x Aa) 3:1 Bi (AaBb x AaBa) 9:3:3:1
Heterozygous x Homozygous Recessive Mono (Aa x aa) = 1:1 Di (AaBa x aabb) = 1:1:1:1
Difference between homology and analogy
Homology: similarity due to shared ancestry Analogy: similarity due to convergent evolution
How can we use these equations to determine what is happening to the gene pool of a population over time?
If genetic frequencies are changing or staying the same
List 2 of the assumptions of the Hardy - Weinberg equilibrium
Individuals do not come into or leave the population No mutations no sexual selection
On the test, page 107, the numbers for the dihybrid cross is given. set up on a table the headings given for the math.
Know to find all the values. 1. add all observed 1st 2. expected are all 100 (in a perfect world) 3. multiply across and don't forget x2 3. di= 3rd line on the probability line. line them up and then your answer is not sign or sign diff
What is required for E. coli to grow? Glow?
LB (Process substance) - to glow, araC
The food for all E.coli bacteria
LB agar
Perfect environment to grow E.coli
Lb agar
Identify the following and environment or genetic: Malaria and Sickle cell anemia
Malaria is environmental Sickle cell anemia is genetic
Tool for transporting small amount of liquid
Micropipette
------------ are the complementary base pairing of nitrogenous bases that form sequences that can be read the same forwards and backwards.
Palindrome
mutations
Random errors in gene replication that lead to a change in the sequence of nucleotides. The source of all genetic diversity.
---------- enzymes have specific active site that recognizes particular nucleotide sequences called recognition sequences.
Restriction enzymes
population genetics
Study of allele frequency distribution and change under the influence of evolutionary processes.
shared derived character
Synapomorphy
Liquid that carries electricity
TAE buffer
Cladogenesis
The process by which sexually reproducing species arise
Taxonomy
The scientific study of how living things are classified
How does size of a DNA band effect how far it moves on the gel
The smaller the DNA fragment, the faster the band will travel.
What is the purpose of preforming a chi square statistical test?
To make a count and see if the observed values are close enough to the perfect expected ratio or are they to far away. We need a statistical test to back up our if they are close enough or not
DNA's backbone is negatively charged and therefore migrates to the positive (red) end of the gel. T/F?
True
The presence of Malaria maintains the sickle cell allele in a gene pool. T/F? explain your answer for credit
True - if it wasn't for the environmental factor of malaria, then the diseased allele for natural selection we would expect to get rid of
p2 = HbA/HbA Homozygous dominant allele q2 = HbS/HbS 2pq HbA/HbS Heterozygous
What is the genotype of villagers with sickle-cell anemia? HbS/HbS
Be able to interpret the results of a ran gel (i.e. banding patterns)
Whatever section lines up exactly with the crime scene is the killed based on the DNA matching
know how to construct a dihybrid punnet square
a 16 square box with either male allele or female allele on the top or side cross multiply -FOIL
population
a group of organisms of the same species populating a given area
What sugar is used to make the gels used in electrophoresis?
agarose
What is a "well"
an indentation in a gel where DNA is inserted
promoter gene turned on by arabinose sugar
araC
Polysaccharide that forms a gel matrix
aragose sugar
gene that provides ampicillin resistance
bla
Form growth by bacteria on agar
colony
The ability for the cell to take up plasmid
competence
Define what it means that a bacterial cell is competent. what was done in this experiment to make the bacteria cells competent.
competence= the cell membrane has holes to allow the plasmid to enter the cell-holes are created by CaCL2
Jill has a mother with O blood and a father withAB. What are the possibilities of blood type that Jill could have? List all that apply.
draw punnet square. ii top, I^A;I^B left side. cross them and it will be A or B blood. -phenotype is the A or B -geno is the I^A or I^B
What happens in population when any of the assumptions of Hardy Weinburg equilibrium aren't met?
evolution
Dominant allele
expresses no matter what the second allele is a Capital letter
altering the genetic makeup of an organisms by giving it extra genetic material through its cell membrane
genetic transformation
stresses the bacteria to encourage taking up plasmid from the environment
heat shock
Heterozygous
homologous chromosomes have different alleles at the same locus hetero means different
Homozygous
homologous chromosomes have the same allele at a given locus Homo means same
What is DNA fingerprinting/profiling and what is one example is can be used?
identifying DNA from a specific individual
What conditions are necessary to have genetic equilibrium?
if things change is called evolution
Dawn has type B blood. Her mom has type O blood. What is/are the possible genotype(s) of Dawn's father? List all that apply.
ii=O Dad could be I^b I^B, I^Bi, I^A I^B Mom is I^Bi
Adaptation
inherited characteristic that increases an organism's chance of survival
Know how to do Chi-square and interpret the results (know how to complete table, formulas are not important)
is commonly used to compare observed results with predictions according to a specific hypothesis
What is an outgroup and how is it used in cladistics?
less closely related to the group being considered that it is to another taxon, is included in the comparison
In this experiment, what structure was given to the + E.coli?
means E.coli was give the plasmid
DNA has a -------- charge and is drawn to the ------- anode of the electrical unit in the electrophoresis.
negative charge; red or +
Recessive allele
only expressed when the second allele is the same
Be able to write out the 2 Hardy-Weinberg equations and be able to use them
p+q=1 p2+2pq+q2=1
genetically engineered plasmid given to the bacteria
pGLO
What plasmid was used in this experiment and what did the genes it contains/code for and do?
pGLO; bla- codes for b-lactamase, araC- promoter gene that turns on the GFP when working with GFP, GFP- when present with arabinose, the GFP will transcribe, translate and produce a green fluorescent protein
Know how to calculate expected numbers from observed
page 105 and 107 has the math of O,E, (O-E), (O-E)2, (O-E)2/E. this gives you the x2 value. based on mon or dihybrid, the numbers will help with expected ratio.
Blood typing- phenotypes, interpret results with antisera, transfusions possibilities, heredity
page 108-109
What is the relationship between sickle cell anemia and malaria? Be able to interpret results from the computer simulation based on this relationship (i.e.study your notes from the computer simulation)
pg 162-173
small circular self-replicating DNA molecule that is separate from the bacterial chromosome
plasmid
primitive character
plesiomorphy
Which plates would you expect to see growth/fluorescence and why?
prediction for -pGLO LB = grows/no glow prediction for -pGLO LB/amp = no grow/ no glow prediction for +pGLO LB/amp = grow/ no glow prediction for +pGLO LB/amp/ara = grow/ glow
What is transformation
process in which one strain of bacteria is changed by a gene or genes from another strain of bacteria
There were a total of 400 corn kernel counted (exactly) by Jack and Jill in the lab last week. 311 of the kernels were purple and 89 were yellow. What are the expected values of (as used on a chi-square analysis) Purple________ Yellow________
purple- 300 yellow- 100 -expected needs to be pefect #s... and total was 400 so 3:1= 300 and 100.
What is the function of restriction enzymes
puts DNA within a particular sequence
If p= 0.25, what is q2? Show your work.
q=0.75 the square both sides q2 = 0.5625 p+q= 1 p2 +2pq+ q2 = 1 (AA+ Aa+ aa=1)=genotypes p= dominant and q= recessive
Know how to use the chart on page 107 in your lab manual to form a conclusion
refer to page 107 and understand how to to be able to tell if it is "not significantly different" or "significantly different"
Phylogeny defined
relatedness among species
What type of enzyme is substrate specific and is necessary for DNA fingerprinting/profiling?
restriction enzymes
What is a palindrome sequence
sequences that are read the same forwards as backwards
what is genetic equilibrium?
situation in which allele frequencies in a population remain the same
Why is sterile technique used in microbiology experiments?
so we don't have contamination of other bacterial cells in our results
Why does DNA respond to electrical current?
the backbone of DNA has a negative charge to it, so it is pulled to the positive end of the chamber.
Determine how far a band of DNA moves in electrophoresis?
the length of the DNA fragment determines how far DNA will travel down a gel
genetic fixation
the loss of all but one possible allele at a gene locus in a population
In this experiment, what structure was given the the "+" E. coli?
these E. coli were given the plasmid pGLO
Small fragments of DNA will travel farther through the gel than larger fragments of DNA when exposed to an electric current. T/F
true
What are some procedures that are part of sterile technique?
washing hands no contamination using new equipment
2 purposes of loading dye
weighs down the DNA and moves at a similar pace to watch the DNA progression of movement of DNA during electrophoresis