2 Credits- half-semester module, 24 March-30 April 2014
Mon & Wed 12:30-1:45 Bio-Pharm 3rd floor conference room
Mon & Wed 2:00-4:00 (first half-hour in conference room, remainder in BioPharm 325).
Chris Simon, Biopharm 305D, 6-4640, <firstname.lastname@example.org>
Graduate Assistant: Russ Meister, Biopharm 325A, <Russell.Meister@uconn.edu>; 6-3947
Readings: will be posted as PDF’s.
Handy reference books: 1) Molecular Systematics, 2nd ed. (Hillis, Moritz & Mable, eds. 1996, Sinauer) especially Chapter 11 by Swofford et al. on Phylogenetic Inference; 2) Molecular Evolution: A phylogenetic Approach (Page & Holmes 1998, Blackwell); 3) Inferring Phylogenies (Felsenstein 2004, Sinauer); The Phylogenetic Handbook (eds. Philippe Lemey, Marco Salemi, and Anne-Mieke Vandamme, 2010).
Lecture Goals: The course will focus on the basics of molecular systematics theory and practice from the point of view of the data. We will explore the ways in which an understanding of processes of evolution of molecular data can help in the construction of evolutionary trees. Lectures will examine some of the most serious problems in evolutionary tree construction: nucleotide bias, alignment, homoplasy, among-site rate variation, taxon sampling, long branches, big trees, heterogeneous rates of evolution among branches, covarion shifts.
Laboratory Goals: Labs will cover basic techniques in molecular systematics from DNA extraction to sequencing, alignment and cloning. This lab will be of interest to both experienced and novice molecular systematists because we will try newly developed kits/techniques and compare them to older ones.
1) For each topic a bibliography will be provided including one focal paper for which the PDF will be posted. Each student will need to turn in a one-page summary of the importance of each focal paper (1 or occasionally 2 papers per week).
2) The week prior to the start of classes you will be given a checklist discussing practical considerations, organization and data checks for molecular systematics. In certain sections you are asked to answer questions and explain how these procedures are modified in your lab.
3) There will be a short "secondary structure alignment assignment" during the semester.
4) Each student will keep a laboratory notebook and hand-in data collected during the course in the form of an alignment and a nexus data file. Various exercises will be performed in laboratory and some will be finished outside of class. These are detailed in the laboratory syllabus.
5) For each Lab, one student will present a 10-15 minute Powerpoint presentation relating to techniques used in that day’s lab. Russ will be available to advise you, but use web searches and try to do as much as possible on your own. These Powerpoint presentations will be posted on the class website so that in the future when you teach a molecular systematics class, they can be used as a starting point to revise and develop lectures of your own.
Final Exam: The final exam will be a take home test in which each student critiques the first draft of a paper submitted to Systematic Biology (submitted in the past but making comments as if it were submitted today). Each student will also compare the submitted version to the published version. The answer key will be the actual review containing reviewers, associate editors, and editor’s comments (with permission of authors, reviewers and editors) and a list of critical points that need to be considered by the authors.
Final Due Dates: Sunday May 4th: Lab project and notebook due. Take Home FINAL EXAM handed out Sunday May 6th. Take home final due Sunday May 11th.
||An introduction to looking at your data: How molecules evolve.
| Reading too big to post, sent out by Chris. How Molecules Evolve Bibliography
|| Data checks at every step. Mechanics of Lab; Explanation of class Tettigades project . Start Nucleospin kit extractions
||The many roles of biological systematics
| Kjer & Honeycutt , Soubrier et al. , Sullivan & Swofford , ASRV Bibliography
|| Mini-presentation: DNA extraction- ultrapure to ultradirty, (phenol-chloroform/CsCl gradients to filters to salting out to chelex, etc.) Russ. Also, Johana gave a small presentation about her work with different extraction methods.
LAB: Chelex extraction. Finish Nucleospin extractions
|| Problems associated combining data, multiple gene histories for single taxa (Species trees and gene trees)
| Bull et al 1993 , Pagel and Mead 2004 , Combining Data & Partitioning Bibliography
|| Before lab, read the introduction to the primer compilation, study the primer comparisons among animals for the COI and COII genes in Simon et al. 1994. And Simon et al. 2006. Mini-presentation: Primer Design- Primer exercise introduction; the beginning of Genious. Russ
LAB: Run extractions on gels. Demonstrate DNA & RNA extraction quantification and the use of the nanodrop. Homework: Troubleshoot and improve “universal” primers for COI and COII in comparison to four complete Tettigades sequences
|| Lecture 4. Choosing partitions, comparing trees
|| Mini-presentation: The Polymerase Chain Reaction- how it works & optimizing reactions. Johana Goyes
LAB: Set-up PCR reaction (mtDNA of Tettigades species, COI barcode, two directions), run gel
|| Lecture 5 Guest Speaker. Paul Frandsen. PDF of Paul Fransden lecture
|| rRNA structure & Alignment Bibliography
|| Mini-presentation: Different methods for cleaning PCR products for sequencing reactions Jimmy Bernot
LAB: Purify PCR products and set-up sequencing reactions
||Lecture 6. Secondary structure & alignment (cont.); Molecular clocks
|| Brandley et al. 2011 , Molecular Clocks Bibliography
SECONDARY STRUCTURE HOMEWORK ASSIGNMENT , associated reading sent by Chris, too large to post
| Mini-presentation: How Big Dye works, chromatograms, and troubleshooting
LAB: Sephadex and put samples on the ABI; Looking at sequences using Sequencher/Geneious, making contigs, blasting sequences in Genbank
||Lecture 7. Long branches, taxon sampling, Felsenstein-zone & anti-felsenstein zone; long branch pruning strategy
|| Long branches, Taxon Sampling, Big Trees Bibliography Swofford et al. 2001 Due April 16th
|| Mini-presentation: - Cloning DNA
LAB: Cloning- Long Lab.
||Lecture 8. Big Trees, Long Branches, & Simulations
|| Mini-presentation: Depositing sequences in GenBank
LAB: PCR clones/Set up sequencing reactions- Long Lab
||Lecture 9: Among Lineage rate variation: nucleotide bias among taxa
|| Nodal Support Readings , Salichos and Rokas 2013 , Betancur et al. 2014 (Comments on Salichos and Rokas 2013) , Summary due April 25th, 2014
|| Mini-presentation: Ancient DNA & Museum DNA protocols
LAB: Sephadex and put clone samples on ABI
||Lecture 10: Tests of topology and problems associated with nodal support
|| Mini-presentation: Numts
LAB: Compare products with those from PCR with DNA vs cloning template and complete mtDNA sequences
||Lecture 11: ALRV: heterotachy, covarion models; long branch problems, taxon sampling, meaning of "basal taxon"
|| Lockhart et al. 2006. Heterotachy and trees (Summarize this one, Wednesday, April 30th) , Lockhart and Steel 2005. A tale of two processes , Gruenheit et al. (Lockhart) 2008. Covarion under changing proportions var sites , Bibliography
|| Mini-presentation: RNA: extraction and what it can be used for
LAB: RNA isolation- Nucleospin RNA Kit
||Lecture 12: Among Lineage rate variation: Covarion evolution: codon models
|| Guest Lecture: Beth Wade, Next Gen sequencing applications, Transcriptomics, Rad Tags, Class Discussion on the implications for modeling data for phylogenetic analysis
|| Lab notebook due. Take home final handed out.
|| Nothing new
|| No Lab
||Final Exam due, emailed to Russ
|| Nothing new
|| You are so done with this class
Final Exam Files
Final Exam Instuctions, Shull et al Manuscript , Shull et al figures, Published Shul et al 2001.
Molecular Systematics Website from 2012 (http://hydrodictyon.eeb.uconn.edu/eebedia/index.php/MolSys2012)