Luria Delbruck Experiment
Goal: To analyze CAN1 sequences from surviving yeast cells after plating on
canavanine (both wild type and mutator strains). We'd like to see which mutations
are non-synonymous.
1- Open Geneious.
2- Import CAN1 sequences and CAN1 reference sequence.
3- Select 5 contigs from each CAN1 sequence group and the reference CAN1
sequence.
4- Choose "Map to reference" from the Align/Assemble folder. CAN1 reference
sequence (because it's longer than each contig) will have already been selected as
reference. Click ok.
5- When looking for mutations, look for majority consensus...Also avoid counting
mutations from sequence regions containing "N", particularly if they don't agree
with other contings.
6- When you find a mutation, or an indel (which you will see as "-"), write down the
number of bp at which it occurs with respect to the reference CAN1 gene (this is
important).
7- Now translate the reference CAN1, and choose frame 1. This is the only frame
with a terminating codon at the end and starter codon in the beginning of the
sequence.
8- To see if the mutation or indel were non-synonymous, first create a copy of the
reference CAN1 gene, turn on editing function, and make the mutation/indel you
observed.
9- Now translate the above sequence (frame 1), and do a protein alignment with the
translated reference CAN1 gene. Do you see a change between these two
sequences?