Lab Prac Assessment

Purification and ANALYSIS of LYSOZYME

Fraction A Fraction B Fraction C Fraction D 26/10/2020 GROUP 12 - Lab Prac Assessment (ONLINE) https://rmit.instructure.com/courses/62987/assignments/512693 1/4 GROUP 12 - Lab Prac Assessment (ONLINE) Due Tuesday by 23:59 Points 10 Submitting a file upload Available 26 Oct at 9:00 - 27 Oct at 23:59 1 day Submit Assignment COURSE NAME: Biochemistry and Molecular Biology 1 COURSE CODE: BIOL1177 ASSIGNMENT TITLE: Lab Practical Exam (ONLINE) DUE DATE: 27-October-2020 Overview of Assignment As previously mentioned, following Stage 4 restrictions, all on-campus learning activities including outstanding practicals in BIOL1177 were paused. Since then we have been rethinking how we can deliver these essential learning activities in different ways so you can continue to study without delay. While Stage 4 restrictions are temporary, RMIT is committed to providing students with certainty so you make informed decisions regarding your study plans. As we progressively transition activities back to campus, we will continue to make decisions based on Government advice and our own campus assessments. Government restrictions may be reinstated at any time dependent on the advice of the Chief Health Officer. As the final assessment for BIOL1177 is now delivered completely online, there will be an alternative assessment for the Laboratory Practical Examination. For those students who attended the laboratory practical examination you will be given the opportunity to undertake this assessment. This means that the mark you received on your face-to-face laboratory practical examination will be superseded by the mark you obtain on this assignment. It is up to you if you wish to do undertake this assignment Assignment Breakdown You will be asked to construct a standard curve, determine appropriate dilutions of an unknown sample, plot a graph (eg. Using Microsoft Excel or any other statistical package), determine the final concentration of the unknown protein and then comment on your results It is recommended you read about the unknown dilution examples presented at the start of the practical manual Taksh Patel Taksh Patel 26/10/2020 GROUP 12 - Lab Prac Assessment (ONLINE) https://rmit.instructure.com/courses/62987/assignments/512693 2/4 BIOL1177 - LAB PRAC EXAM (ONLINE) You will be provided with a Proforma which provides details on the practical assessment and space is provided for you to complete tables, calculations, draw your graph and comment on your results. Please read the instructions on the proforma carefully DO NOT MODIFY the proforma TYPE your answers in the space provided You must conform to the page limit. If you go over the page limit you will be penalised This assessment is worth 10% of your overall mark This assessment will be open on Monday 26th of October and 9AM and will close on Tuesday 27th of October at MIDNIGHT Access your assignment here: BIOL1177 2010 Laboratory Practical Assessment ONLINE 73 FINAL.docx Learning Outcomes 1. Explain the central role of biochemistry and molecular biology within the health and biomedical disciplines. 2. Describe and explain the principles and fundamental concepts of biochemistry and molecular biology with a focus on analytical techniques, proteins, enzymes, Vitamins and Trace Elements, DNA and RNA metabolism and gene regulation. 4. Problem solve, validate and interpret results of biochemical investigations. RMIT Electronic Submission of work for assessment I declare that in submitting all work for this assessment I have read, understood and agree to the content and expectations of the assessment declaration (https://www.rmit.edu.au/students/student-essentials/assessment-and- exams/assessment/assessment-declaration) . Please see additional info on pdf: Accessible Powerpoints.pdf   https://rmit.instructure.com/courses/62987/files/15392224/download?wrap=1 https://www.rmit.edu.au/students/student-essentials/assessment-and-exams/assessment/assessment-declaration https://rmit.instructure.com/courses/62987/files/9864302/download?wrap=1 26/10/2020 GROUP 12 - Lab Prac Assessment (ONLINE) https://rmit.instructure.com/courses/62987/assignments/512693 3/4 Criteria Ratings Pts 0.5 pts 0.5 pts 0.5 pts 1.0 pts 0.5 pts 3.0 pts 0.5 pts Aim 2-3 Sentences 0.5 Pts Exemplary Aims: A clear and concise description of what the experiment is trying to achieve should be presented. The following keywords should be mentioned: calibrations/standard curve, quantify, Bradford, unknown albumin. 0.0 Pts No marks Aims: A clear and concise description of what the experiment is trying to achieve is unclear or missing. The following keywords should be mentioned: calibrations/standard curve, quantify, Bradford, unknown albumin are missing. PART 1. Point 5. Example Calculation 0.5 Pts Exemplary The formula C1V1=C2V2 has correctly been applied (formula and all working out) for a selected tube from Table 1. 0.0 Pts No marks The formula C1V1=C2V2 has been applied (formula and all working out) for a selected tube from Table 1 but some or the majority of details are incorrect or missing. TABLE 1 0.5 Pts Exemplary Table 1 is complete and correct (volumes of standard, volume of water, final volume, and final concentration and corrected absorbance) has correctly been presented. An appropriate table caption has been included. 0.0 Pts No marks Table 1 is incomplete with either (volumes of standard, volume of water, final volume, and final concentration and corrected absorbance) missing or incomplete. An appropriate table caption has not been included. PART 2. Point 3. 1.0 Pts Exemplary Appropriate and feasible dilution factors have correctly been applied (including working out) with a detailed explanation of why these are required. 0.5 Pts Accomplished Appropriate and feasible dilution factors have been applied (including working out) with an explanation of why these are required but some minor errors are present. 0.0 Pts No marks Dilution factors have been applied with a limited or no explanation of why these are required. TABLE 2 0.5 Pts Exemplary Table 2 is complete and correct (volumes of unknown, dilution factors, dilution volumes, corrected absorbance, unknown protein concentration, and mean protein concentration) has correctly been presented. An appropriate table caption has been included. 0.0 Pts No marks Table 2 is incomplete with either (volumes of unknown, dilution factors, dilution volumes, corrected absorbance, unknown protein concentration, and mean protein concentration) missing or incomplete. An appropriate table caption has not been included. PART 4. Bradford Standard Curve 3.0 Pts Exemplary Graph (Bradford standard curve) has clearly labelled axes and units, duplicate values plotted, line of best fit and a appropriate figure caption has been included. 2.0 Pts Accomplished Graph (Bradford standard curve) has clearly labelled axes and units, duplicate values plotted, line of best fit and a appropriate figure caption has been included but some minor errors are present. 1.0 Pts Developing Graph (Bradford standard curve) has clearly labelled axes and units, duplicate values plotted, line of best fit and a appropriate figure caption has been included but major errors are present. 0.0 Pts No marks Graph (Bradford standard curve) has been incorrectly graphed labels and axes, duplicate values, line of best fit and a appropriate figure caption are missing or incomplete. PART 4. Point 2. 0.5 Pts Exemplary The concentrations of the unknown protein samples (Neat, Dilution 1 and 2) (Tubes 13 - 18) have been drawn and labelled on the graph. 0.0 Pts No marks The concentrations of the unknown protein samples (Neat, Dilution 1 and 2) (Tubes 13 - 18) have incorrectly or not been drawn and labelled on the graph. 26/10/2020 GROUP 12 - Lab Prac Assessment (ONLINE) https://rmit.instructure.com/courses/62987/assignments/512693 4/4 Total points: 10.0 Criteria Ratings Pts 2.0 pts 1.0 pts 0.5 pts PART 4. Point 3 2.0 Pts Exemplary The final protein concentration (including calculations) is correct. A detailed discussion as to which dilution was used is well justified. 1.5 Pts Accomplished The final protein concentration (including calculations) has some minor errors presented. A detailed discussion as to which dilution was used is reasonably justified. 1.0 Pts Developing The final protein concentration (including calculations) has some major errors presented. A detailed discussion as to which dilution was used is satisfactorily justified. 0.0 Pts No marks The final protein concentration (including calculations) is incomplete or incorrect. A detailed discussion as to which dilution was used is missing or incomplete.PART 4. Point 4 1.0 Pts Exemplary A discussion on the concept of linearity and associated errors have been interpreted correctly, demonstrating an excellent understanding of results conveyed. 0.5 Pts Accomplished A discussion on the concept of linearity and associated errors have been presented, demonstrating an adequate understanding of results conveyed. 0.25 Pts Developing A discussion on the concept of linearity and associated errors have been presented but major improvments are required. 0.0 Pts No marks Incomplete or incorrect interpretation of the concept of linearity and associated errors, indicating a complete lack of understanding.Conclusion 0.5 Pts Exemplary A single paragraph which clearly and concisely describes the outcome(s) of this experiment has been presented. 0.0 Pts No marks A single paragraph which clearly and concisely describes the outcome(s) of this experiment is missing or incomplete. Quantitation of human albumin in a patient’s plasma sample Student Name: _________________ GROUP NUMBER: ______________________ Student Number: ______________________ Demonstrator: _________________________ Aim:________________________________________________________________________________________ __________________________________________________________________________________________________________________________________________________________________________________________ (0.5 mark) Part 1. Prepare your standard curve. 1. You are provided with a standard albumin protein solution (200 ug/mL). 2. Create a 6-point standard curve from 0 ug/mL – 50 ug/mL. Each standard should be performed in duplicate (tubes 1 - 12). (Absorbance values are provided below). 3. The final volume in each tube should be 0.5 mL. 4. Determine and record the final concentration and volume for each tube in Table 1below (include caption). 5. Provide an example calculation for one of your tubes by applying the formula C1V1=C2V2 below. Ensure you show all working out. _____________________________________________________________________________________________________________________________________________________________________________ (0.5 mark) ______________________________________________________________________________________________________________________________________________________________________________________________ Table 1. ________________________________________________________________________________ Tube 1 2 3 4 5 6 Standard albumin (200 µg/mL), mL H2O, µL Final volume/tube, µL Final [albumin] (µg/mL) Absorbance (595 nm) 0.402 0.424 0.568 0.560 0.661 0.658 Absorbance - Blank Tube 7 8 9 10 11 12 Standard albumin (200 µg/mL), mL H2O, µL Final volume/tube, µL Final [albumin] (µg/mL) Absorbance (595 nm) 0.771 0.770 0.877 0.881 0.980 0.955 Absorbance - Blank (0.5 mark) Part 2. Prepare your unknown patient’s plasma sample. 1. You are provided with an unknown protein solution containing albumin in the range 0 – 150 ug/mL. 2. You will need to perform 2 serial dilutions of the unknown albumin sample. You will need to prepare between 1 - 2 mL of the first dilution to be able to perform the next serial dilution. 3. Determine and record the appropriate dilution factors for both dilutions 1 and 2 in Table 2 below. Explain in detail why your unknown needs be diluted. ____________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________ ___________________________________________________________________________________________ (1.0 mark) 4. Set up microfuge