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23 Cards in this Set

  • Front
  • Back
what is the mutation we are working with
what does the mutation affect?
the second virescent mutant that was isolated in arabidopsis thaliana and is a nuclear recessive mutation that affects chloroplast development
what is the mutant phenotype
leaves become green much more slowly that the leaves of wildtype plants and the leaves are never as fully green as wildtype levels of chlorophyll.
what does RFLP stand for
restriction fragment length polymorphism
how many RFLPs do we use
what is an RFLP
single base pair changes that occur in the chromosomes of the species under study
what are the two different ecotypes of arabidopsis
Landsberg (L) and columbia (C)
what is an ecotype
geographically separated populations of arabidopsis that still have the ability to be crossed together and produce normal progeny. due to nucleotide changes over time, different ecotypes have slightly different genomic DNA sequences
what are microsatellite markers
repetitive DNA based on dinucleotide repeats
what were the phenotypes of the parental generation that was crossed
vir2 columbia mutant x wildtype landsberg
what are the genotypes of the parental generation
vir2vir2 CC x vir2+vir2+ LL
what is the phenotype of the F1 generation
a combination of L and C with one member of each pair of homologous chromosomes deried from the L parent and the other from the C parent
what is the genotype of the F1 generation
vir2+vir2 CL
how many pairs of chromosomes does arabidopsis have. how many are sex chromosomes
5. 0.
how many of the F2 generation have the vir2 phenotype
why are the chromosomes of the F2 generation a mixture of C and L
crossing over occurs during meisois when the two C and L homologues form a tetrad. the result is that the chromosome I that enters the gametes is part L and part C
if an F2 plant has the vir2 phenotype what is its genotype? why?
vir2vir2. bc vir2 is recessive. since vir2 originally came from the C parent, than a plant that is vir2vir2 must have C DNA in the region of the vir2 gene on both homologues
it must be CC in the region of the chromosome wher the vir2 gene is located
what is the gel visualized with
ethidium bromide
what are the ingredients to a PCR reaction
template DNA, deoxyribonucleotides (dNTPs), buffer, foward and reverse primers, taq DNA polymerase, and water
What was the total volume of each reaction
20 ul
What are the steps to a PCR reaction and what are the temperatures?
1. denature dna strands at 94
2. 92
3. anneal primers at 54
4. extend with taq polymerase @ 72
5. hold at 20

repeat 30 cycles
explain the difference between RFLPs and microsatellite markers
cutting RLFPs with certain restriction enxymes will give us one pattern of cleavage for columbia and a different pattern for landsberg
microsatellites will have different sizes between landsberg and columbia
what RF value determines a linkage
an RF<0.5 means that they are linked and that the two alleles are on the same chromosome
otherwise they are far apart or on separate chromosomes