Read Background information on your Series 2 Forward Genetics Project at BISC219/F11:_Gene_Mapping_Info. Submit statement of Series 2 experimental question & goals. Outline (or create flow diagram) our experimental process including : Mutant Phenotype description compared to WT; Linkage Analysis; Mapping the Mutation to a chromosome and to a particular chromosomal location; Complementation Analysis; Defining the gene sequence and protein change through DNA sequence analysis; & Bioinformatics to find homologues and to determine if the mutation is new or previously characterized. Write 1-2 page narrative summary of the investigation BISC219/F11: Assignment_Series2_Outline_Summary.
Diagram the expected results (genotype and phenotype)for the crosses and self-fertilization progeny in the Linkage Testing work. Use only part of the template provided at Media:Series 2 Template for Crosses.ppt (Note that you will use and submit only through Linkage 2. (The Mapping part of the template will be used next week). Include a narrative description of this experimental design that you have drawn out on the template and summarize what you will be looking for in the results that will allow determination of the linkage group to which your mutated gene of interest belongs. Assignment information and rubric found at BISC219/F11: Assignment_Series2_Linkage Testing Crosses
Diagram the expected results (genotype and phenotype)for the crosses and self-fertilization progeny in the Mapping work that you will do over the next several weeks by completing the templates provided. Make sure you highlight the progeny that will be used to continue each step of the process. Assignment information and rubric found at: BISC219/F11: Assignment_Series2_Mapping Crosses.
Analyze your mutant gene's DNA sequence using the instructions that you can download from Lab 6. Construct a Results and Discussion section, WITH A FIGURE, for this analysis. Assignment information and rubric found at: BISC219/F11: Assignment_Series2_DNA Sequencing
Convert all Series3 Protocols, performed so far, as Materials & Methods. This Assignment is described at BISC219/F11: Assignment_Series3_ Materials and Methods. Due at the beginning of Lab 9 ; Read the original 1998 paper by Fire and Mello published in Nature391: 806-811 (Feb.19 1998) at doi:10.1038/35888 ; Do your own library search to find and read a recent review article about RNAi in C. elegans. Also read the 2002 Simmer et al. paper in Current Biology12:1317-1319 explaining the rrf-3 strain that we will use for our RNAi work at doi:10.1016/S0960-9822(02)01041-2 . These articles provide background information on how the mechanism of RNAi was worked out experimentally and will be helpful to you when you write the introduction section of your Series3 paper. Also read a published journal article by Green et al. (2009) Impact of Cigarette Smoke Exposure on Innate Immunity: A Caenorhabditis elegans Model. PLoS ONE 4(8): e6860, found at doi:10.1371/journal.pone.0006860. This study, like yours, uses reverse genetics to investigate gene function. Be prepared to discuss this paper in Lab 9.
Write a draft introduction section (including properly formatted Literature Cited page) of your next paper on our Reverse Genetics Project. Refer to BISC219/F11:Resources Guide to Scientific Writing.
Nov. 24 for all students
Construct figures/tables with properly formatted legends to illustrate the main findings of the RNAi part of your Series3:Reverse Genetics project. These figures are a draft of those that will be part of the results section of the research report you will write for Series3.