Mapping vir 2 mutation in Arabidopsis thaliana 1. How do you determine the genot
ID: 3166981 • Letter: M
Question
Mapping vir 2 mutation in Arabidopsis thaliana1. How do you determine the genotype of an individual plant at each marker locus?
2. How would you decide what percentage of aragose tonuse to resolve the DNA fragments in this Vir2 lab? How would you select an appropriate molecular weight marker to compare the fragments to?
3. Once a marker with close linkage is found, what steps would be taken to identify the mutant gene?
Mapping vir 2 mutation in Arabidopsis thaliana
1. How do you determine the genotype of an individual plant at each marker locus?
2. How would you decide what percentage of aragose tonuse to resolve the DNA fragments in this Vir2 lab? How would you select an appropriate molecular weight marker to compare the fragments to?
3. Once a marker with close linkage is found, what steps would be taken to identify the mutant gene?
1. How do you determine the genotype of an individual plant at each marker locus?
2. How would you decide what percentage of aragose tonuse to resolve the DNA fragments in this Vir2 lab? How would you select an appropriate molecular weight marker to compare the fragments to?
3. Once a marker with close linkage is found, what steps would be taken to identify the mutant gene?
Explanation / Answer
1. SSLP markers can be differentiated using PCR and gel electrophoresis. SNPs that create restriction sites (CAPS marker) can be differentiated using PCR, restriction digestion and gel electrophoresis.
2. For SSLP markers, use native PAGE gel to resolve DNA bands if the size difference is less than 10 bp. If the size difference is more than 10 bp, agarose gel can be used.
For CAPS markers, agarose gel can be used to visualize restriction digestion pattern.
3. Once a mutation is fine mapped to a given region, candidate gene approach should be followed. We can select a few candidate genes and analyze their function by studying previous reports about their function. If any gene is reported to exhibit similar function, we can sequence that particular gene.
We can simply get the entire region sequenced to find out the location of the mutation.
If any mutant plants are available for the genes present in the given genomic interval, we can get those mutants and cross them with the unknown mutant to establish allelic status.