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My Genetic Profile Worksheet Human genetic characteristics and phenotypes Characteristic Dominant Phenotype Dimples Present (D) Hairline Widow’s peak (W) Tongue rolling Roller (R) Thumb placement with folded Left thumb on top (P) hands Recessive Phenotype Absent (d) Straight (w) Non-roller (r) Right thumb on top (p) Parent #1 Phenotype Genotype Dimpled Dd Straight ww hairline Tongue roller Rr Left thumb on PP top Parent #2 Phenotype Genotype Dimpled Dd Widow’s peak Ww Non- roller Right thumb on top rr pp Child (F1) Phenotype Genotype Grandchild (F2) Phenotype Genotype Pedigree Chart for a SINGLE characteristic Parent #4 _____ Parent #3 _____ Parent #2 _____ Parent #1 ____ Child #2 _____ Child #1 _____ Grandchild _____ STAO 2011 – The Promise and Pitfalls of Genetics (SBI3U) Session #1019 www.facebook.com/nelsonschoolsciencek12 Presenter: Doug Fraser What are the chances? In this investigation you will model a cross between two heterozygous individuals. You will then determine the genotype and phenotype ratios of your model F1 generation. In addition, you will investigate the role that sample size and probability play in producing a 25:50:25 ratio in the F1 generation. Equipment and Materials: two small opaque bags containing 40 beads each (20 blue beads and 20 red beads) Note: Red beads are the dominant allele, R, for red flowered, and the blue beads are the recessive allele, r, for blue flowered. Together, the two pouches represent the parents of the P1 generation. Without looking, draw one bead from bag 1 (P1) and one bead from bag 2 (P2). This represents the joining of two gametes to form a new individual. Record the “genotype” of this offspring. Return the “gametes” to their original bags. Repeat for a total of 20 times. Offspring Genotypes 1 2 11 12 3 4 5 6 7 8 9 10 13 14 15 16 17 18 19 20 Expected Genotype ratio: ______ RR : ______ Rr: ______ rr Expected Phenotype ratio: ______ Red : ______ Blue Actual Genotype ratio: ______ RR : ______ Rr: ______ rr Actual Phenotype ratio: ______ Red : ______ Blue How did your results compare to the expected ratio? How did the total class results compare to the expected ratio? STAO 2011 – The Promise and Pitfalls of Genetics (SBI3U) Session #1019 www.facebook.com/nelsonschoolsciencek12 Presenter: Doug Fraser A Velcro Model for Microarrays A Velcro analogy model (VAM) can be used to effectively introduce and model the principles of microarray technology. Prior to conducting the activity the teacher introduces the basic theory/concepts behind the use of microarray technology as follows: DNA/RNA basics • When a gene is active in a cell the DNA sequence of the gene is transcribed into an mRNA molecule. • The mRNA delivers this genetic instruction into the cytoplasm where it is used to make a protein or part of a protein (a polypeptide) • RNA is very similar to DNA in that for every base in the DNA gene sequence, the cell assembles a matching RNA base. • Therefore the mRNA molecule will have a set of bases that are complimentary to the original DNA strand. • NOTE: Geneticists are able to use mRNA from a cell to make a complimentary copy of DNA (cDNA) that will match up with original DNA in the gene. Original DNA strand G C A T G G A A T C C... Complimentary DNA C G T A C C T T A G G... (made using mRNA) Microarray basics • Microarrays contain hundreds or thousands of clusters of single-stranded DNA molecules attached to specific locations on a glass slide • Each cluster consists of identical copies of DNA that corresponds to a specific gene. So one microarray will have clusters of DNA corresponding to 100’s or 1000’s of different genes. o Note that these are not “entire” genes but large fragments of the coding portions of the genes – enough to be unique to each gene. • Each DNA cluster will be strongly attracted to any cDNA made from complimentary mRNA strands. For example: DNA strands with the base sequence TTCAGGCAG will be attracted to any cDNA strands with the sequence AAGTCCGTC. In other words each DNA cluster will be attracted to cDNA that were made using mRNA sequences that were transcribed from the same gene. • The microarray is exposed to cDNA made using ALL the mRNA being produced by a particular group of cells. The different cDNAs will become “stuck” to their matching DNA clusters. • One can then determine which genes were active in the cells simply by observing which clusters of DNA have cDNAs attached to them! • By comparing the microarray results for different cells you can compare gene activity in each cell! STAO 2011 – The Promise and Pitfalls of Genetics (SBI3U) Session #1019 www.facebook.com/nelsonschoolsciencek12 Presenter: Doug Fraser Liver tumor tissue Normal liver tissue Diseased pancreas cells Gene Membrane protein #1 Respiration protein #1 Unknown #1 Unknown #2 Membrane protein #2 Milk protein Ribosomal protein Unknown #3 Insulin Respiration protein #2 Hemoglobin Glucagon Normal pancreas cells Microarray Observations and Analysis: Questions: 1) Compare the activity of genes in normal pancreas cells with those of diseased pancreas cells. What might this suggest about the possible cause of the disease and effect that this disease may have on the cells normal activity? 2) Compare the activity of genes in normal liver tissues with those of liver tumor tissue. What might this suggest about the possible cause of the cancer and effect that this may have on the tissues normal activity? 3) Compare the activity of genes in normal pancreas and normal liver cells. Why aren’t the same genes active in all healthy human cells? 4) Which genes were active in all cells? What might this suggest about the roles and importance of these genes? 5) Which genes were not active in any of the cells? Where in the body would you expect to find these genes active? STAO 2011 – The Promise and Pitfalls of Genetics (SBI3U) Session #1019 www.facebook.com/nelsonschoolsciencek12 Presenter: Doug Fraser The VAM Microarray: Each pair should have the identical microarray Velcro (hooks) board and two envelopes with sets of cDNA strands (Velcro loops) – each envelope representing the cDNA from a given tissue/cell sample. Make board out of “foam core” board and use adhesive-backed Velcro. Microarray Board Layout: Membrane protein #1 (long) AATCGGCGATCT Unknown #2 (medium) TTGCGGAAT Ribosomal protein (long) CGGATTATCCGG Respiration protein #2 (medium) ACTGTTACC • • • Respiration protein 1 (medium) CGCCTTATT Membrane protein #2 (short) TATAGG Unknown #3 (short) CGTAAA Hemoglobin (long) AATGAGGGAATT Unknown #1 (medium) CCCTAGGAG Milk protein (long) GATCCGGACCGT Insulin (short) CGTAAT Glucagon (short) TACACC Make DNA (Velcro) strips in three different lengths Write base codes and protein identification above DNA Velcro strips that are on the board Attach labels with base codes on the back of each separate sample DNA strip to be placed in envelopes So for each “board” you will need four long, four medium, and four short pieces of Velcro. Plus labels that include the gene “name” and the base sequence The following is assuming you will have 8 microarray “boards” and one group at each board – with two envelopes per board as follows: Groups 1,2,3 - Normal Pancreas and Diseased Pancreas cells Group 4,5,6 – Normal Liver tissue and Liver Tumor tissue Groups 7,8 – Normal Pancreas and Normal Liver tissue cells. Envelope contents: Normal pancreas cells:Mem1, resp1, unknown 1, mem2, ribo, insulin, resp2, glucagon Diseased pancreas cells:Mem1, resp1, mem2, ribo, resp2, glucagon Normal liver tissue: Mem1, resp1, unknown 2, ribo, resp2 Liver tumor tissue: Mem1, resp1, unknown 2, ribo, unknown 3, resp2 Compliment codes for cDNA velcro in envelopes Membrane protein 1 AATCGGCGATCT TTAGCCGCTAGA Unknown #2 TTGCGGAAT AACGCCTTA Ribosome protein CGGATTATCCGG GCCTAATAGGCC Respiration protein #2 ACTGTTACC TGACAATGG Respiration protein 1 CGCCTTATT GCGGAATAA Membrane protein #2 TATAGG ATATCC Unknown #3 CGTAAA GCATTT Hemoglobin AATGAGGGAATT Not needed Unknown #1 CCCTAGGAG GGGATCCTC Milk protein GATCCGGACCGT Not needed Insulin CGTAAT GCATTA Glucagon TACACC ATGTGG STAO 2011 – The Promise and Pitfalls of Genetics (SBI3U) Session #1019 www.facebook.com/nelsonschoolsciencek12 Presenter: Doug Fraser