Genetics Lab report

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There are TWO parts.

Part 1 does not require a formal lab report. Just give your answers and the results of the cross. For part 2, a type written lab report is needed. Use the handout to help you put together your report. You MUST have a cover page.

All details are in the uploaded pdf file. Please read it carefully and follow it exactly as required.

Refer to the handout provided as Principles of DNA Lab (1).

Lab report format:

Scientific laboratory reports or any other APA style paper written at a collegiate

level have a standard format.

The following are standard for APA papers:

1 inch margins on all sides

12-point font and the font style should be either Arial, or Times Roman

Some type of Header (Running Head:)

Numbered pages except the Title Page

Paragraphs indented 1-tab space

DOUBLE-SPACED throughout the paper including Title Page and Reference Page.

Scientific laboratory reports have a separate Title Page and a separate Reference page.scientific reports have HEADINGS

on each section of the report.

These headings are:

Introduction

Materials& Methods

Results

Discussion

TITLE:

The first page of the lab report is the TITLE page. Do NOT number the Title page unless instructed to do so.

The title of the lab report should NOT be too general but should reflect more specifically about the experiment. For example: “Fingerprinting” is too general for the title but instead, use something like: “Fingerprinting as an Effective Forensic Tool in Solving Murders.” The title should be informative; it should not be “cute.” A Running head is needed as a Header on all pages including the title page.

The Running Head should appear as Running Head: SHORT TITLE OF PAPER IN ALL CAPS (but not in bold).


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Department of Biological and Environmental Sciences BIO L320 – GENETICS LAB Dr. Jacqueline Jones Laboratory 5 – Mendelian Genetics – Principle of Independent Assortment lab and Principles of DNA Sequencing Principle of Independent Assortment: In this section of today’s lab, we will conduct two exercises related to Mendel’s Principle of Independent Assortment (Investigation 1): Exercise 1. The first experiment is a pen and paper exercise based on the dihybrid cross for Drosophila ebony body X vestigial wing described on page 11 (old book) & page 13 (new book), part A, number 1. You need to calculate the expected results for the F1 phenotypes, and F2 phenotypes and frequencies as described on page 11 (old book) & page 15 (new book), part A, number 2. In addition, you need to show (a) the genotype of the F1 flies resulting from this cross, (b) the haploid gametes that the F1 individuals would produce, and (c) produce a Punnett square for the F2 generation of this dihybrid cross, showing all genotypes and phenotypes. You will have to write this part up on a sheet of paper, DO NOT use the lines at the bottom of page 11 (old book) & page 15 (new book). (25 points) Principles of DNA Sequencing: Exercise 2. The purpose of this exercise is to provide detailed instruction on the practice of DNA sequencing. You will be provided an actual autoradiograph (an exposed sheet of x-ray film) of a DNA sequencing gel for analysis. The sequence deduced from the autoradiogram will differ from a wild type sequence by a single nucleotide(s). Refer to the handout provided on Blackboard. This difference represents an actual mutation in the DNA molecule. You are expected to identify the location(s) of the mutated nucleotide(s). You can also work in groups to read the sequence and compare results to assure accuracy. Make sure you read the handout because you will see this information again on your midterm. (25 points) Part 1 does not require a formal lab report. Just give your answers and the results of the cross. For part 2, a type written lab report is needed. Use the handout to help you put together your report. You MUST have a cover page. Total = 50 points Pages needed to be copied from the lab book: 11 and 13 (old book); 13 and 15 (new book). Also Print out other Document for Exercise 2. 1 Principles of DNA Sequencing Part II Background Information The purpose of this exercise is to provide detailed instruction on the practice of DNA sequencing. The student is provided an actual autoradiograph (an exposed sheet of x-ray film) of a DNA sequencing gel for analysis. The sequence deduced from the autoradio­ gram will differ from a wild type sequence by a single nucleotide. This difference rep­ resents an actual mutation in the DNA molecule. The students are expected to identify the location of the mutated nucleotide. Students should also work in groups to read the sequence and compare results to assure accuracy. Rapid analysis of DNA sequence was developed during the 1970's from research groups in the United States and England. Since its early days, these methods have been refined and automated. There are two basic approaches to DNA sequence analysis. One involves a set of organic chemical reactions with the DNA bases. The other uses an enzymatic process. The chemical method is tedious and labor-intensive, whereas the enzymatic approach, which is often called the dideoxy method, is quite fast. The autoradiographs you have been given as part of this experiment, are the result of the enzymatic procedure which uses the Kienow fragment of the E.coli DNA polymerase I to make a DNA copy of the region to be sequenced. More commonly, a DNA polymerase known as sequenase or equivalent is used for DNA sequencing. I A specialized cloning vehicle constructed from an E. co/ivirus, called M13, facilitates rapid DNA sequence analysis. This virus contains a polylinker, which is a short region of DNA, about 57 base pairs, containing several unique restriction sites. Segments of DNA to be sequenced are inserted into the polylinker region using standard cloning procedures (Figure 1}. The M13 virus contains a single-stranded circular genome with about 7200 nucleotides. The virus will infect E. coli strain JM101 cells which contain a fertility factor. These cells are F·, and male. The virus infects by attachment to the sex pilus. Shortly after infection, the viral DNA will become double­ stranded. It is this form which serves as a template for produc­ tion of single-stranded DNA progeny. The DNA associates with the viral proteins to form mature virus and the virus exits the cell by budding; the cell is not lysed. For cloning and sequenc­ ing purposes, double-stranded DNA to be sequenced is inserted into the M13 polylinker region of the DNA intermediate, and then transformed into competent JM101 E.coli cells. The trans­ formed cells will begin to produce progeny virus. polylinker region .--::w..�---- To sequence DNA which has been inserted into the polylinker region of M13 single­ stranded DNA is prepared from viral plaques. In this experiment, a short 17-base synthet­ ic single-stranded DNA is allowed to hybridize (form a base pairing} with a unique site in M13 adjacent to the polylinker. I II site of hybridization of sequencing primer Figure 1 Experiment Overview and Procedures EXPERIMENT OBJECTIVE: The objective of this experiment is to develop an understanding of DNA sequencing and analysis. This is a dry lab which contains autoradiographs from an actual DNA sequencing experiment. I EXPERIMENTAL PROCEDURE: 1. Obtain the sample autoradiograph and place it on a light box to enhance visualiza­tion. 2. The sequencing reactions have all been loaded in order: G-A-T-C. 3. Begin analysis of the DNA sequence at the bottom of the autoradiograph with the circled band, which is an A. 4. Identify the location of the mutant nucleotides. What was the mutation? Is there more than one mutation? 5. To help you start, the sequence begin with an 5' A. I • I I Drosophila and Maize Experiments in Genetics: Monohybrid and Dihybrid Crosses 11 1. What F2 phenotypic ratio do you expect to obtain? 2. What F2 genotypic ratio is expected? 3. Which trait is dominant in your experiment? 4. How do you know? B. Maize Crosses The inheritance of the extensive genetic variability in maize also obeys Mendelian principles; both seedling and aleurone (endosperm) characters lend themselves to classroom use. Figure 1.7 shows a flat of F2 seedlings segregating for a recessive mutant allele for albinism (absence of chlorophyll). The classical Mendelian ratio of 3:1 is expected. Similarly, Figures 1.8 and 1.9 show ears of F2 kernels of corn that are segregating in a ratio of 3colored : 1colorless aleurone typical of a monohybrid cross, or in a ratio of 1colored : 1colorless aleurone typical of a monohybrid testcross. Can you use the photos to confirm these expectations? Because maize has a relatively long life cycle (3 months or more to complete), you probably will not be able to conduct actual experimental matings with it. Your instructor might provide you with ears of genetic corn or flats of F2 seedlings and request that you determine the number of individual seeds or plants having the various phenotypes and then interpret the data in terms of Mendelian principles. For example, suppose you counted 40 green and 12 albino seedlings in an F2 population. How would you interpret these data? FIGURE 1.7 Flat of F2 corn seedlings segregating for a recessive albino mutation. The classical Mendelian ratio of 3:1 is expected. Drosophila and Maine Experiments in Genetics Monohybrid and Dlhybrid Crosses 13 TABLE 1.3 Record of F, Data for Maize Indosperm Trait Number of Kernels Observed Phenotypes Number of Kernels Expected Number Genotypes of Kernels Phenotypes will vary depending on the trait studied Ears bearing F, kernels can be purchased from a number of biological supply companies. Without removing the kernels from the ears, you can count the number of kernels in the different phenotypes, record data in Table 1.3, and then formulate the hypotheses to explain the data collected. Two pos- sible illustrations of endosperm traits follow F2 I SUSHI Parents 1 SUSH X SUSU starchy sweet (smooth) (wrinkled) 2. CC X сс pigmented nonpigmented (purple) (white) F, Susu starchy (smooth) 3 Su- starchy (smooth) 3 C- pigmented (purple) Sweet (wrinkled) : lcc nonpigmented (white) pigmented (purple) A host of other endosperm mutants have been researched. Many are able for classroom labo- ratory investigation. Examples include Wx (starchy) versus wx (waxy), Y (yellow) versus y (white), and Pr (purple) versus pr (red) endosperm. Franks (1980, 1981) and Neuffer et al. (1997) provide further information and excellent color photographs of useful maize mutants. VII. INDEPENDENT ASSORTMENT A. Dihybrid Cross with Drosophila Under the direction of the instructor, prepare a mating between flies carrying homozygous mutant genes in the second and third chromosomes. Produce Fi and F2 generations. Several crosses are appropriate for this experiment. Although one such cross will be suggested here, your instructor may decide to use alternative crosses. 1. The gene for vestigial wings (vg) is located on the second chromosome, and the gene for ebony body (@) color is located on the third chromosome. Matings may be made between flies from these two stocks. Be certain to use virgin female flies for this cross. Reciprocal crosses, that is, vestigial female (ugog; c'e') x ebony male cug'ug'; ee) and ebony female X vestigial male, may be made if time and equipment permit. After about 8 days, remove and discard the parent flies. Record all data relative to this experiment in Table 1.4. У 17 AG TT GGTC (GG 104T GG A7 а! 2 AGC 77 4G LTC64 7CaAcac AT icar 7 (CCC (04ATTG7AATA 1 2 CCCCAGGAATTGTAATA 28
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Running head: DNA SEQUENCING

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DNA Sequencing
June 6th, 2018

DNASEQUENCING

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Introduction

The concept of DNA sequencing is one of the most important scientific discovery of
recent times. DNA sequencing by definition refers to the process of determining the exact order
of nucleotides within a given DNA molecule. The process comprises of various technologies
and methods that are employed for the determination of the order of the four bases (adenine,
cytosine, quanine and thymine) of any given DNA strand. The invention of rapid methods of
DNA sequencing has led to...


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Excellent resource! Really helped me get the gist of things.

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