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Unformatted text preview: BSC 2010L BSC POPULATION GENETICS II & POPULATION INVERTEBRATE DIVERSITY I INVERTEBRATE 1. In this lab, we will: 1. In A. Complete the Polymerase Chain Complete Reaction (PCR) exercise. Reaction B. Collect data for the Hardy-Weinberg Collect analysis. analysis. C. Begin our study of invertebrate diversity. PCR Exercise - Second Week Procedures Procedures • Pipette 7 µ l into a well in the gel. • Be sure to record the loading order of the samples. • Each gel should also have at least one lane with the 7µ l of the Pgem ladder. • We will run the gels at 100 V for 30 min. • Then we will photograph the gels on the UV transilluminator. • Finally, you will score the gels. – The band with the insert will run slower than the band without the insert. Is our population in Hardy-Weinberg Equilibrium for the Alu insert? Equilibrium Alu Insert primer Alu insert (300bp) primer 400 bp Alu insert missing primer primer 100 bp How to read the final gels: How • These data are used to complete the Hardy-Weinberg Equilibrium data sheet(pp. 183-184, 10 pts.) This is due at the end of lab. • In addition, you must write an abstract of this PCR exercise (10 pts.). This will be due at the next lab. Assumptions of Hardy-Weinberg Equilibrium Equilibrium Conditions for genetic equilibrium: a. b. c. d. e. random mating very large population = no genetic drift no selection: all genotypes are equally viable no mutation no migration = no gene flow H/W Equation H/W equation predicts the number of individuals of each genotype if the population is in equilibrium – Need to know the allele frequencies in the population – Plug those allele frequencies into the equation to get the expected genotype frequencies for a population in equilibrium – Compare those predicted genotype frequencies to the observed genotype frequencies – Remember: gene frequencies of a particular generation depend upon the gene frequencies of the previous generation and not upon the genotype frequencies – Students frequently use genotype and allele frequency interchangeably in lab reports, so stress the difference How do we get allele frequencies? • If you know the genotype of all the individuals in the population, then you can calculate the allele frequencies • For example: – – – – – 25 individuals are AA 17 are AB 30 are BB What is the allele frequency of A? What is the allele frequency of B? Calculating Allele Frequencies Genotype AA AB BB Totals: # of individuals 25 17 30 ___ #A __ __ __ ∑ ___ #B __ __ __ ∑ ___ A allele frequency = p = ∑A / (# of individuals x 2) B allele frequency = q = ∑B / (# of individuals x 2) To relate allele frequencies to genotypic frequencies for 2 alleles (A & B): 1. p = proportion of the A allele q = proportion of the B allele 2. since there are only two alleles: p + q = 1 – and so: p = (1-q); q = (1-p) 3. Hardy Weinberg Equation: p2 + 2pq + q2 = 1 4. p2 = probability (proportion) of homozygous ‘AA’ 2pq = probability heterozygous ‘AB’ q2 = probability homozygous ‘BB’ To test if a population is in HardyTo Weinberg equilibrium Compare observed genotypes with expected genotypes 1. To get the expected number of each genotype: – p2 x # individuals in sample = expected number of AA individuals. – 2pq x # individuals = expected number of AB individuals. – q2 x # individuals = expected number of BB individuals. 2. Compare the expected genotypic numbers obtained above with those observed from the gel electrophoresis. – Use a chi-square test • Note that in a chi square test you must compare numbers of individuals, not frequencies. – Degrees of freedom are equal to 1. (If p + q =1, once you know either p or q, you know the other.) Review of chi-square test 2 2 Observed AA AB BB Expected o-e (o-e) (o-e) /e Chi-square = Σ Invertebrate Diversity – Invertebrate Part I Part PORIFERA THRU ANNELIDA PORIFERA Principal Biological Concepts to Emphasize: to A. Classification B. Coloniality vs. multicellularity. C. Specialization of form and function of cells. C. Specialization D. Organization of cells into tissues of specific structure and D. function. function. E. Organization of tissues into organs. E. Organization F. Organization of organs into organ systems. F. Organization G. Trends in body plan. G. Trends H. Organ systems and function. H. Organ Developments in body organization organization A. Tissue organization - diploblastic vs. triploblastic. A. Tissue 1. Mesoglea is not a tissue. 2. Outer ectoderm or epidermis, inner endoderm or Outer gastrodermis. gastrodermis. 3. Triploblastic have mesoderm. B. Radial versus bilateral symmetry B. Radial C. Cephalization. C. Cephalization. D. Coelom development and function. D. Coelom 1. Concentration of sensory apparati at the anterior end of Concentration the body. the 2. An adaptive response to bilateral symmetry and linear An mobility. mobility. 1. Body cavity isolating internal organs from body wall. 2. Protects organs. 3. Acts as a hydrostatic skeleton. Structure and Function 1. support 1. 2. movement 3. circulation 4. respiration 5. nervous response 6. ingestion 7. digestion 8. excretion 9. reproduction a. sexual b. asexual c. cycles d. ploidy 10. osmoregulation • In small organisms, many functions occur by In diffusion. diffusion. ORGAN SYSTEM Int eg um ent ar y PRIMARY FUNCTIONS Cr e at e s a ba r r ier b e t w ee n inte r n al st r uctu re s and th e o ut sid e ­ pr o t ec t io n ag ains t in j u r y , d esicc at io n, inf ect io n Mo v em ent , loc o m o t io n Supp or t, p ro te c ti o n o f inte r n al st r uctu re s Gas e xch an ge , t ake in ox y ge n, r e le ase CO2 Pro c essin g of f oo d ; nu t r iti o n ­ ing est io n, di ge st io n, ab so r p ti o n , eg est io n ( eli m in at io n) Exc r e t io n o f m et ab ol ic (n it r o ge n o us) wa st es; o sm ore gu lati o n ­ wa t er and io n ba lanc e in bo d y Mo v em ent o f m at er ials w it hin / th ro u g h o ut b o d y Co lle c ti o n o f ly mph , d ef ens e ag ains t inf ect io ns and c an c e ro us c ells Re p r o du c ti o n , p ro duct io n o f ga me t e s, d ev elo pm ent o f zy g ot e and emb ryo Coor din at e ac t iv iti es, p er c eiv e and r e sp o nd t o st imu li Chem ic al c oor din at io n w it hin b o d y Muscu lar Ske le t al Re spi ra t or y Dig est iv e Exc r e to r y Cir c ul at ory L y mph at ic / im mun e Re p r o du c ti v e Nerv o u s End o c r in e Functions of structures detailed in the manual will not always be included in the text. You are still required to know the function. If you do not know the function, you should ask me or look it up. ask Classification Classification A. Review taxonomic hierarchy - Domain, A. kingdom, phylum (division), class, order, family, genus, species. order, B. 3 domains B. domains 1. Bacteria 2. Archaea 3. Eukarya Eukarya a. "Protists" – At least 5 kingdoms "Protists" b. Kingdom ANIMALIA b. Kingdom c. Kingdom FUNGI c. Kingdom d. Kingdom PLANTAE d. Kingdom Kingdom ANIMALIA - KNOW TAXONOMY TO CLASS. CLASS Will cover chapters 17 and 18. Will Systematics have changed Systematics somewhat with new information from molecular studies. studies. Phylogeny based of body plan grades grades 1. Parazoa - no true tissues - sponges 1. Parazoa 2. Eumetazoa - true tissues 2. Eumetazoa A. Radiata - radial symmetry, Radiata diploblastic - Cnidaria diploblastic B. Bilateria - bilateral symmetry, Bilateria triploblastic triploblastic 1. Acoelomates - Platyhelminthes 1. Acoelomates 2. Pseudocolomates - Nematoda 2. Pseudocolomates 3. Coelomates 3. Coelomates - Protostomes - Mollusca, Protostomes Annelida, Arthropoda Annelida, Phylogeny based on molecular studies molecular 1. Parazoa - no true tissues - Porifera 1. Parazoa 2. Eumetazoa - true tissues 2. Eumetazoa A. Radiata - radial symmetry, diploblastic Radiata Cnidaria Cnidaria B. Bilateria - bilateral symmetry, triploblastic 1. Protostomes 1. Protostomes - Lophotrochozoa - Platyhelminthes, Platyhelminthes Mollusca, and Annelida Mollusca, - Ecdysozoa - Nematoda and Nematoda Arthropoda Arthropoda Animal phylogeny based on sequencing of SSU-rRNA Animal Comparing the molecular based and grade-based trees of animal phylogeny Comparing Phyla CALCAREA and SILICEA - sponges SILICEA A. Sponges (the Parazoa) are distinct A. from all other animals (the Eumetazoa). Eumetazoa 1. Lack tissues 1. Are either asymmetrical or radially symmetrical symmetrical A choanoflagellate colony choanoflagellate Sponges Sponges Anatomy of a sponge Anatomy Phylum CNIDARIA A. Radial body symmetry (the Radiata) Radiata B. Polyps and medusae in life cycle B. Polyps C. Diploblastic - two tissues layers C. Diploblastic 1. Ectoderm 2. Gastroderm 3. Also have mesoglea - not a tissue D. Classification: D. Classification: 1. class Hydrozoa- polyps dominant 2. class Scyphozoa - medusae dominant 3. class Anthozoa - medusae absent Polyp and medusa forms of cnidarians Polyp Medusa Medusa Purple striped jelly, Pelagia panopyra Pelagia Sea anemones Sea Coral polyps Coral The life cycle of the hydrozoan Obelia Obelia Remainder of animals have bilateral symmetry (the Bilateria) Bilateria A. Cephalization A. Cephalization B. Triploblastic - 3 tissue or germ layers B. Triploblastic 1. Ectoderm 2. Mesoderm 3. Endoderm Coelom development Coelom a. Acoelomates a. Acoelomates b. Pseudocoelomates b. Pseudocoelomates c. Coelomates c. Coelomates Phylum PLATYHELMINTHES - ACOELOMATE ACOELOMATE A. class Turbellaria - planarians A. class B. class Trematoda - flukes B. C. class Cestoda - tapeworms - internal parasites parasites Anatomy of a planarian Anatomy Protonephridia: the flame-bulb system of a planarian Protonephridia: Anatomy of a parasitic flatworm Anatomy Anatomy of a tapeworm Anatomy Phylum NEMATODA roundworms, hookworms PSEUDOCOELOMATE PSEUDOCOELOMATE A. Ascaris A. B. Turbatrix Turbatrix C. Trichinella - DEMO Trichinella Nematode, C. elegans C. Parasite nematode, Trichinella spiralis Trichinella Phylum ANNELIDA segmented worms COELOMATE COELOMATE A. class Polychaeta - mostly marine worms A. class B. class Oligochaeta - e.g. earthworm B. e.g. C. class Hirudinea - leeches C. class Annelids, the segmented worms: Polychaete (left), feather-duster worm (middle), leech (right) Annelids, External anatomy of an earthworm External Invertebrate Scavenger Hunt • Complete the Inverts I scavenger hunt data sheet (pp. 205-206, 5 pts.) • Find answers on the display cards associated with the display specimens. • Turn this in before you leave lab. • Work alone on this. Inverts Study Aid Inverts Prelab exercises Prelab A. Complete the Earthworm dissection and the Crayfish A. dissection prelabs before coming to the next lab. dissection ...
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