Course Title: Apply electrophoretic techniques

Part B: Course Detail

Teaching Period: Term1 2014

Course Code: ONPS5662C

Course Title: Apply electrophoretic techniques

School: 155T Vocational Health and Sciences

Campus: City Campus

Program: C5282 - Diploma of Laboratory Technology (Biotechnology)

Course Contact: Amberlee Mitton

Course Contact Phone: +61 3 9925 8053

Course Contact Email: amber.mitton@rmit.edu.au


Name and Contact Details of All Other Relevant Staff

Nominal Hours: 100

Regardless of the mode of delivery, represent a guide to the relative teaching time and student effort required to successfully achieve a particular competency/module. This may include not only scheduled classes or workplace visits but also the amount of effort required to undertake, evaluate and complete all assessment requirements, including any non-classroom activities.

Pre-requisites and Co-requisites

MSL973002A  - Prepare working solutions OR MSL974001A - Prepare, standardise and use solutions
MSL974003A   - Perform chemical tests and procedures.

Course Description

This unit of competency covers the ability to analyse samples using electrophoretic techniques. The unit also includes establishing client needs for routine and non-routine samples, optimising enterprise procedures and instruments for specific samples, obtaining valid and reliable data and reporting test results. Personnel are required to recognise atypical test data/results and troubleshoot common analytical procedure and equipment problems.


National Codes, Titles, Elements and Performance Criteria

National Element Code & Title:

MSL975008A Apply electrophoretic techniques

Element:

1. Establish client needs and schedule analysis

Performance Criteria:

1.1. Liaise with client or sample provider to determine client needs and sample history
1.2. Record sample description, compare with specification and record and report discrepancies
1.3. Identify non-routine samples and the possible need to vary enterprise procedures
1.4. Seek advice from supervisor about any proposed variations and document all approved changes
1.5. Schedule analysis using enterprise procedures

Element:

2.    Prepare samples and standards

Performance Criteria:

2.1. Obtain a representative analytical portion of the laboratory sample
2.2. Prepare sample in accordance with testing requirements
2.3. Prepare validation checks for analytical portion

Element:

3.    Set up and optimise instrument

Performance Criteria:

3.1. Perform pre-use and safety checks in accordance with enterprise procedures
3.2. Start up and condition the instrument using enterprise procedures
3.3. Optimise instrumental parameters to suit sample and test requirements
3.4. Check calibration status of instrument and perform calibration using specified standards and procedures, if applicable

Element:

4.    Perform analysis

Performance Criteria:

4.1. Measure analyte response for standards, validation checks and samples
4.2. Conduct sufficient measurements to obtain reliable data
4.3. Return instruments to standby or shutdown condition as required

Element:

5.    Process and analyse data

Performance Criteria:

5.1. Confirm data is the result of valid measurements
5.2. Perform required calculations and ensure results are consistent with standards or estimations and expectations
5.3. Record results with the appropriate accuracy, precision and units
5.4. Analyse trends in data and/or results and report out of specification or atypical results promptly to appropriate personnel
5.5. Troubleshoot analytical procedure or equipment problems which have led to atypical data or results

Element:

6.    Maintain a safe work environment

Performance Criteria:

6.1. Identify risks, hazards, safety equipment and control measures associated with sample handling, preparation and analytical method
6.2. Use personal protective equipment and safety procedures specified for test method and materials to be tested
6.3. Minimise the generation of wastes and environmental impacts
6.4. Ensure the safe disposal of laboratory wastes
6.5. Clean, care for and store equipment and consumables in accordance with enterprise procedures

Element:

7.    Maintain laboratory records

Performance Criteria:

7.1. Enter approved data and results into laboratory information management system
7.2. Maintain equipment logs in accordance with enterprise procedures
7.3. Maintain security, integrity and traceability of samples and documentation
7.4. Communicate results to appropriate personnel


Learning Outcomes



Details of Learning Activities

Lab classes and Record keeping
Lectures/Workshops
Team meetings
Presentations
Exams
Reports and Assignments
Online theory and quizzes
Research
Fieldtrips


Teaching Schedule


Program is subject to change
FOR LONG EXPERIMENTS THEORY & PRAC CLASSES ARE MERGED TOGETHER


Part 1 - DNA

8/4 8 Exam week


(week
of)   
 week  THEORY
(at home)
 PRAC 4 Hours Assessment
10/2 1  Intro to Electrophoresis
**On campus class**
 
   No lab class this week
 
Quiz
(in class)
17/2 2 Visualising DNA:
 3 useful stains, Molecular weight markers,
Gel concentrations 
Agarose Gels I: Frankenstein Gels - Separating DNA with different % gels.

Preparing solutions -  Agarose gel electrophoresis (calculations)
Quiz
24/2 3 Quantifying DNA using agarose electrophoresis and fluorimeter methods.  Agarose Gels II How much & how pure is my DNA? 
  
Quiz
3/3 4 Which allele(s) of Alu
is in your DNA? How can you tell from the electrophoresis results?
 Electrophorese Mol Gen
Alu PCR products gel electrophoresis
Quiz
10/3 5  Electrophoresis of RNA (Northern Blotting); Electrophoresis of Genomic DNAs (Pulsed field gel electrophoresis, Southern Blotting)   1. Pipetting Workshop (Eppendorf*)
*Refresher - using pipettes
*How to calibrate pipettes 

2. Prac report 1 annotating gel photos
Quiz
Prac report 1 due – do and submit in class
17/3 6 Intro to DNA sequencing & Capillary Electrophoresis        DNA Sequencing*
(AGRF Sequencing facility visit)
Quiz
24/3 7 Revision (at home)    
 30/3  8  Exam week   Exam 1
 7/4  9  Protein Electrophoresis Types of gels: agarose, cellulose acetate,
SDS-PAGE & MW calculations.
 1. Prepare solutions for protein agarose gel & SDS- PAGE.

2. Agarose Gel Electrophoresis of Serum Proteins

(Helena Labs*)
 Quiz
 14/4  10 Intro to SDS-PAGE,
Calc size of proteins using SDS-PAGE
 Protein Agarose Electrophoresis of Serum Haemoglobins
(Helena Labs*)
 Quiz
    29th Mar – 4th Apr Student vacation    
 29/4 11  Dotblot Immunoassay, Western transfer & Westernblot Immunoassay Dotblot
 6/5 12 quiz SDS-PAGE of Albumin and ovalbumin.
Prac Report 2
  

Quiz

Prac Report 2
SDS-PAGE of Albumin
 

 13/5 13 quiz  Record results from last week
SDS-PAGE of Fish Proteins.
Quiz
 20/5 14 quiz Western Transfer and Blot of Fish Proteins. (BioRad*)
 

Quiz

(prac report due)
Logbooks due

 27/5 15  REVISION  No lab class - revision at home  
 3/6 16  Exam Week    Exam 2 Exam 2
 10/6   Alternative assessments (Special consideration only)   Alt Exam if required
     * To be confirmed    


Learning Resources

Prescribed Texts

Lab Manual - Term 1 (DNA)
Lab Manual - Term 2 (Protein)
Both Lab manuals are available from the course Blackboard. Please print the manual in time for the week 1 class. Print the whole manual at the start of the semester - bring it to class each week so you have the instructions for each experiment. The Term 2 Manual will be available towards the beginning of term 2.

Resources available on Blackboard for this course


References


Other Resources


Overview of Assessment

Assessments for this course typically consist of:
 

  • Class activities
  • Reports
  • Presentations
  • Exams
  • Online Quizzes
  • Logbook records of experiments


Assessment Tasks

 

Assessment 1. Theory
a. Weekly Quizzes 20% (best 10 grades count @ 2% each) There is a quiz most weeks
b. 2 Theory Exams 25% + 25% = 50% Weeks 8 and 16

Assessment 2. Prac Reports
1: Agarose Electrophoresis of DNA 15% Week 5
2: SDS-PAGE of Albumin 15%  Week 12

Assessment 3. Logbook
Satisfactory completion required
(competent records kept for at least 80% of lab classes)
Records must be signed off by the teacher each week. 
Submit in week 14
 

Total     100%        


Assessment Matrix

Other Information

Important information about assessment

To pass the course you need to pass, on average, each type of assessment
(exams, reports, quizzes, logbooks).

For example, there are two exams, so you need to have an average of 50% to pass AND you also need to have passed the other assessments as well. You can’t make up marks from one type of assessment to another (eg pass the tests but fail the prac reports).

If you miss a prac class you must provide a copy of your medical certificate or other evidence (court appearance, funeral notice, statutory declaration) to the teacher.

Extension of time for submission of assessable work:
A student may apply for an extension of up to 7 days from the original due date
They must lodge the application form (available on the web) at least the day before the due date
The application is lodged with the School Admin Office on Level 6, Bdg 51
Late work that is submitted without an application for an extension will not be corrected, and therefore will receive a zero grade.
Students requiring longer extensions must apply for Special Consideration (form available on the Web)

For missed assessments such as exams- you (& your doctor if you are sick) must fill out a special consideration form. This form must be lodged at the HUB or online with supporting evidence (eg medical certificate), prior to, or within, 48 hours of the scheduled time of examination.

Do each week’s Quiz before the Lab class to help you prepare.

Each quiz is available from the time it is released until the start of prac class time.
Network or technical difficulties will not be accepted as a valid excuse for not having done the quiz
 

 


Logbook Assessment
The logbook is a record of your experiments, and your activities in class.
Your logbook must be signed off after completion each week by both the student AND the teacher.
The teacher will also signoff your logbook on the class sheet as a central record.

The central record is the evidence record that is required to show that your logbook has been signed off by you, the student, and the teacher.

IT IS THE STUDENT’S RESPONSIBILITY TO GET THEIR WORK SIGNED OFF EVERY WEEK.
(If you miss the class you do not need to get the sheet signed off – the teacher will record this as an absence). If a logbook is not signed off, that week’s grade will be NYC* unless you were absent.

*NYC = not yet competent

When is a lab performance considered “competent’ (ie when can a logbook be signed off)?
Logbooks will only be signed off when the student has followed and completed a method adequately, and is therefore considered competent for that experiment.

This is evidenced by:

THE LOGBOOK WILL ONLY BE SIGNED OFF BY THE TEACHER AFTER THE FOLLOWING:

1. Your performance during the lab class is satisfactory
2. Show date of analysis/results at top of page at start of experimental record in logbook,
3. Attach loose pages to logbook securely (use tape or staples),
4. Include raw and analysed results in logbook as appropriate to particular method (see lab manual)
5. You have cleaned up lab work area and equipment adequately
6. You have signed off logbook entry once record of results and the clean up are completed

IDEALLY THE LOGBOOK IS SIGNED OFF ON THE DAY OF THE EXPERIMENT.
If this is not possible due to time constraints (the teacher will let you know when this is the case), you can get the logbook signed off WITHIN 2 WEEKS of doing the experiment. Logbooks will not be signed off after this time.

At least 80% of labs must be recorded so that sufficient skills development can occur.

When will a logbook not be signed off?

A logbook entry won’t be signed off (ie it is considered ‘not yet competent’ for an experiment if:

• The Student doesn’t attend class long enough to complete the experiment
• The Student doesn’t finish the procedure in a reasonable time
(nominally by the time class finishes – excluding when there are logistic issues that mean few/no one in the class finishes).
• The student doesn’t contribute to the group effort adequately
• Results are a debacle – several errors/extensive lack of following SOP (method).
• The experiment is not conducted in a safe manner
• The lab area and equipment are not adequately cleaned up

In the case of a not yet competent result, note that there is no opportunity to repeat that particular experiment due to scheduling of classes. In this case, to achieve competency other experiments during the course should be done to a ‘competent’ level and the student should show a good amount of improvement as semester goes along as evidenced by observations in class by the teacher.

Booklist

1. A logbook is required for this course to record all experimental details. The logbook must be brought to class each week and signed off by the teacher.
The book needs to be a bound NON-SPIRAL exercise book.

Completion of experimental records in the logbook trains students how to keep good records that can be used as “proof of discovery”. These logbooks form legal documents that can be used to prove discovery for Intellectual Property & patenting purposes & thus form an important part of the electrophoresis course. Because pages can easily be ripped out of SPIRAL bound books,
SPIRAL bound books ARE NOT ACCEPTABLE.

All loose pages should be attached to the logbook as soon as possible. This is so that important experimental records do not get lost.

The logbook is to be submitted at the end of the semester – see the assessment guide.


2. Resources available from Blackboard including:
• Theory notes and worksheets
• Videos and diagrams that show each electrophoretic technique used in class
• Links to internet resources
• Online quizzes

Time Allowance for this course
The 100 nominal hours for this unit are allocated as follows:

Activity Breakdown (hours)
theory at home (1hr*7)+(1hr*5)+5 (week1) + 4 (week7) = 17
online weekly quiz 10 quizzes * 1.5hr = 15
revision at home 2 * 5hrs = 10
exams 1&2 2 * 2hr = 4
prac (7*4hr) +(3*5hr) = 43
fieldtrip 4hr = 4
report write ups 2 * 3.5hrs = 7

Total Hours = 100

 


 

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