BL 415 2011 Lab test 2 study guide for second exam: ~1.5 hour lab test 250 pts
Understand the basic principles of the experiments, the basic procedures and the data analysis.
Reread the prelabs, the experiments, lab reports and review answers to questions;
Review any new terms and what they mean, especially those related to lecture concepts.
There will be gels, example data to analyze – Chi square formula will NOT be provided, but the P value
significance table will.
Know the general safety precautions for the lab, disposal of petri plates, sharps, used glass pipets, etc.
Review different stains, dyes we used,
Review equipment and its function (vortex, minicentrifuges, etc)
G1 - sizes of 3 pipettors, which pipet to use for which volumes
Microliter vs. milliliter and liter – how many ul in a ml? etc.
V1C1 = V2C2 relationship of diluting solutions:
2 ml of 50% glucose in total of 200 ml = 0.5% final concentration
2 ul of 1 ug/ul DNA = 2 ug total DNA
Temperatures used for different procedures – and are centigrade
G2 model organisms and scientific names, sketch of each organism
– why are they experimental models?
Arabidopsis mutants observed
Recall which are prokaryotes and which eukaryotes
Virtual Fly lab. What did we do and what was found?
Monohybrid, dihybrid crosses, sex-linked, recombination
Testing data fit to monohybrid, dihybrid by chi-square
Testing data fit to hypothesis of no linkage -> chi-square won’t give good fit of data, so
conclude linkage and calculate % recombination (don’t do chi-square on the % recomb)
Mapping genes by three-point cross; order or genes, genetic distance
Why 10,000 flies instead of 1000?
G4 Alu PV92 allele PCR of human polymorphic marker.
What is an Alu element? Why so named?
What is PCR, how did you isolate DNA, and what solutions in isolation and PCR reactions?
PV92 is a dimorphism, and not a polymorphism – why?
What is seen on the gel – (- vs. + allele); also recall Semi-log plot and calculation of sizes of PCR
fragments from markers
Bioinformatics – what is a BLAST search and why do we do them? How did you calculate the
size of the PCR product from BLAST search
Hardy-Weinberg calculations of allele frequencies – be able to analyze gels and interpret data
From numbers of organisms, calculate observed genotype frequencies, observed allele freq.,
expected frequencies, numbers of organisms to calculate chi-squared fit to hypothesis
S4 (III), S5 (II), S6 (I, III) Human genetics
Barr bodies and sex chromatin content of people
In situ hybridization,
Karyotypes and diseases with wrong numbers of sex chromosomes, other chromosomes
G5 Real Drosophila crosses.
What were the crosses we did, which genes , phenotypes, which are sex-linked?
What was the basic procedure – P, F1 and F2 generations?
What was expected for the results of F1 and F2 progeny?
Why were virgin females so important for crossing parents for the F1?
Be able to analyze F2 progeny data by chi-square; see if it fits 3:1 ratio Mendelian
Be able to analyze F2 progeny for sex-link ratios – consider the reciprocal crosses
G6 Yeast transformation
What are principles of yeast shuttle vector – what selectable markers
What is a yeast library – how many clones to screen the genome if insert certain size?
The principle of the Sau3A partial fragments ligated into BamHI site –
Recognition by restriction enzymes; partial digest
What is principle of yeast transformation – what chemicals were used
How were yeast transformants selected? What selectable media
How were transformants screened for having LEU or HIS genes?
How did you determine number of viable yeast? Consider serial dilution calculations
Calculation of transformation frequency; then calculation of number of HIS+/URA+
Consider variation in number of viable yeast in the class – and determining the mean
G7 Worm siRNA – what is siRNA and what is function in normal cell?
Diagram the basic design for this experiment, and expected results.
What were the negative control plates, and the experimental test plate?
Why can worms be fed E. coli and develop altered phenotypes?
What is the principle of siRNA – would you see a change in the genomic DNA of worms
If you had isolated that genomic DNA?
Why was it important to add young worms to the Petri plates vs. big old worms?,
or to only examine young worms?
Wormbase database
S7 NCBI genomic mining – general principles of that site.
PubMed
Chromsome viewer
S8. Population genetics – Hardy-Weinberg equilibrium and its basic assumptions
Cattle – all three genotypes were visible, be able to calculate allele and genotype frequencies
N and M and NM blood groups – all genotypes visible - calculate
For the taste papers, which allele was dominant?
How was the frequency of T or t allele in the population determined?
How could you calculate the equilibrium genotype frequencies?
Blood type – what are the three alleles, and what determines different blood groups.
Diagram the tests for blood types – what is in the anti-A serum, for ex.
You will not need to calculate H-W equilibrium for these blood groups.