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Evidence Guide: MSL975014A - Perform molecular biology tests and procedures

Student: __________________________________________________

Signature: _________________________________________________

Tips for gathering evidence to demonstrate your skills

The important thing to remember when gathering evidence is that the more evidence the better - that is, the more evidence you gather to demonstrate your skills, the more confident an assessor can be that you have learned the skills not just at one point in time, but are continuing to apply and develop those skills (as opposed to just learning for the test!). Furthermore, one piece of evidence that you collect will not usualy demonstrate all the required criteria for a unit of competency, whereas multiple overlapping pieces of evidence will usually do the trick!

From the Wiki University

 

MSL975014A - Perform molecular biology tests and procedures

What evidence can you provide to prove your understanding of each of the following citeria?

Interpret and schedule test requirements

  1. Review test request to identify samples to be tested, test method and equipment/instruments involved
  2. Identify hazards and enterprise control measures associated with the sample, preparation methods, reagents and/or equipment
Review test request to identify samples to be tested, test method and equipment/instruments involved

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Identify hazards and enterprise control measures associated with the sample, preparation methods, reagents and/or equipment

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Receive and handle samples

  1. Log and label samples according to enterprise procedures
  2. Record sample description, compare with specification and note and report discrepancies
  3. Store samples in accordance with enterprise and test method requirements
  4. Maintain chain of custody, traceable to the worker, for all samples
Log and label samples according to enterprise procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Record sample description, compare with specification and note and report discrepancies

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Store samples in accordance with enterprise and test method requirements

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain chain of custody, traceable to the worker, for all samples

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare equipment and reagents

  1. Set up equipment/instrumentation in accordance with test method requirements and perform pre-use and safety checks
  2. Select and collect reagents in accordance with test method requirements
  3. Prepare and label reagents in accordance with test method requirements
Set up equipment/instrumentation in accordance with test method requirements and perform pre-use and safety checks

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Select and collect reagents in accordance with test method requirements

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prepare and label reagents in accordance with test method requirements

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Extract, verify and manipulate biomolecules

  1. Produce/extract biomolecules from samples using appropriate isolation methods
  2. Prevent contamination of samples by unwanted biomolecules
  3. Recognise the presence of common inhibitors of biomolecular reactions and take corrective action
  4. Quantify and qualify biomolecular yields from purified extractions
  5. Use appropriate techniques to prepare and test a range of biomolecular samples
  6. Use controls and reference standards to confirm the integrity of biomolecular sample preparation and procedures
Produce/extract biomolecules from samples using appropriate isolation methods

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Prevent contamination of samples by unwanted biomolecules

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Recognise the presence of common inhibitors of biomolecular reactions and take corrective action

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Quantify and qualify biomolecular yields from purified extractions

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Use appropriate techniques to prepare and test a range of biomolecular samples

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Use controls and reference standards to confirm the integrity of biomolecular sample preparation and procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Process data

  1. Record test data noting atypical observations
  2. Ensure results are consistent with reference standards and expectations
  3. Record and report results in accordance with test methods
  4. Interpret trends in data and/or results and report out of specification or atypical results promptly to appropriate personnel
  5. Troubleshoot basic procedure, reagent or equipment problems which have led to atypical data or results
Record test data noting atypical observations

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure results are consistent with reference standards and expectations

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Record and report results in accordance with test methods

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Interpret trends in data and/or results and report out of specification or atypical results promptly to appropriate personnel

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Troubleshoot basic procedure, reagent or equipment problems which have led to atypical data or results

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain a safe work environment

  1. Use established safe work practices and personal protective equipment to ensure personal safety and that of other laboratory personnel
  2. Minimise the generation of wastes
  3. Ensure the safe disposal of wastes, including hazardous wastes and tested samples
  4. Clean, care for and store equipment and reagents
Use established safe work practices and personal protective equipment to ensure personal safety and that of other laboratory personnel

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Minimise the generation of wastes

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Ensure the safe disposal of wastes, including hazardous wastes and tested samples

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Clean, care for and store equipment and reagents

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Report and communicate results

  1. Record approved data into enterprise system
  2. Keep accurate, traceable work records to protect the enterprise's intellectual property rights
  3. Maintain confidentiality and security of enterprise information and laboratory data
  4. Maintain equipment logs in accordance with enterprise procedures
Record approved data into enterprise system

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Keep accurate, traceable work records to protect the enterprise's intellectual property rights

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain confidentiality and security of enterprise information and laboratory data

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Maintain equipment logs in accordance with enterprise procedures

Completed
Date:

Teacher:		 Evidence:

 

 

 

 

 

 

 

Assessed

Teacher: ___________________________________ Date: _________

Signature: ________________________________________________

Comments:

 

 

 

 

 

 

 

 

Instructions to Assessors

Evidence Guide

The Evidence Guide provides advice on assessment and must be read in conjunction with the performance criteria, required skills and knowledge, range statement and the Assessment Guidelines for the Training Package.

Overview of assessment

Critical aspects for assessment and evidence required to demonstrate competency in this unit

Assessors should ensure that candidates can:

conduct work practices in an ethical and professional manner and in accordance with relevant legislation, regulation and codes of practice

maintain security, integrity, traceability and identity of samples, sub-samples and work records

obtain purified biomolecules from samples

prevent/minimise DNA/RNA contamination

perform tests/procedures, such as PCR, ligation and restriction enzyme digestion with appropriate controls

follow enterprise safety standards, procedures and practices

follow enterprise procedures and test methods consistently and accurately

operate test equipment to enterprise standards and/or manufacturer's specification

identify atypical results as out of normal range or an artefact

trace and source obvious causes of artefacts

communicate identified problems to a supervisor

record and communicate results as per enterprise procedures.

Context of and specific resources for assessment

This unit of competency is to be assessed in the workplace or simulated workplace environment.

This unit of competency may be assessed with:

MSL974008A Capture and manage scientific images

MSL975001A Perform microbiological tests

MSL975008A Apply electrophoretic techniques

MSL975009A Apply routine chromatographic techniques

MSL975013A Perform tissue and cell culture techniques

MSL975020A Apply routine spectrometric techniques.

Resources may include:

laboratory equipped with appropriate test equipment/instruments, standards and reagents

enterprise procedures, standard methods, manuals and supplier documentation.

Method of assessment

The following assessment methods are suggested:

review of test records and workplace documentation completed by the candidate

review of results obtained by the candidate over a period of time to ensure accurate and consistent results are obtained within required timelines

observation of candidate isolating, purifying, verifying and manipulating biomolecules

oral or written questioning

feedback from peers and supervisors.

In all cases, practical assessment should be supported by questions to assess underpinning knowledge and those aspects of competency which are difficult to assess directly.

Where applicable, reasonable adjustment must be made to work environments and training situations to accommodate ethnicity, age, gender, demographics and disability.

Access must be provided to appropriate learning and/or assessment support when required.

The language, literacy and numeracy demands of assessment should not be greater than those required to undertake the unit of competency in a work like environment.

This competency in practice

Industry representatives have provided the case studies below to illustrate the practical application of this unit of competency and to show its relevance in a workplace setting.

Biomedical

As part of a diagnostic service to verify progenitor status of livestock, a technician is required to extract DNA from a blood sample, perform the PCR to amplify micro-satellite DNA and prepare the sample for DNA electrophoresis and fragment size analysis. The technician provides documentation to meet evidentiary standards. The technician understands the implications of the tests for the client and is careful to ensure the sample can be traced from the source, that no contamination takes place and that the results are kept confidential.

Food processing

A meat export company has commissioned a study of the effectiveness of the introduction of a 'cold-chain' process to a client country. The company requires rapid results. As part of the monitoring team, a technician is required to perform routine testing of surface swabs of meat samples for bacterial contamination using a PCR analytic technique. Although the tests are quite routine, the technician pays close attention to all aspects of the work as the consequences of invalid results would be severe for the company and laboratory. The technician also keeps comprehensive work records and maintains strict confidentiality.

Required Skills and Knowledge

Required skills

Required skills include:

conducting work practices in an ethical and professional manner and in accordance with relevant legislation, regulation and codes of practice

maintaining security, integrity, traceability and identity of samples, sub-samples and work records

performing molecular biology tests and procedures

following enterprise safety standards, procedures and practices

identifying atypical results as out of normal range or an artefact

tracing and sourcing obvious causes of artefacts

communicating identified problems to a supervisor

recording results according to enterprise procedures

Required knowledge

Required knowledge includes:

hazards and risks in molecular biology laboratories

common biotechnology terms

molecular biology principles and concepts underpinning tests/procedures, such as:

DNA and RNA structure and function

protein structure and function

relationship between chemical and physical properties of nucleic acids and proteins and the techniques used for sampling, preparation and testing

replication

transcription, translation and gene regulation

relationship between structure, organisation and function of biomolecules to the storage of information in cells, chromatin, circular and linear chromosomes, RNA, genes and plasmids

molecular genetics (molecular nature, organisation and function of genes)

molecular mechanisms of DNA mutation and variation

DNA transfer in prokaryotes (transformation, conjugation and transduction)

restriction enzyme and ligase structure, nomenclature, function, specificity and stability, and cohesive versus blunt ends

ethical issues associated with biotechnology, such as:

use of animals for research

genetic modification of organisms and food

use of gene therapy, cloning and stem cells

in vitro fertilisation

forensic testing of populations

importance of commercial confidentiality, protection of intellectual property and patents

genetic screening of humans

sex determination and parentage testing of embryos/humans

importance and appropriate use of validation methods, controls and certified reference materials

enterprise and/or legal traceability requirements

relevant health, safety and environment requirements

Range Statement

The range statement relates to the unit of competency as a whole. It allows for different work environments and situations that may affect performance. Bold italicised wording, if used in the performance criteria, is detailed below. Essential operating conditions that may be present with training and assessment (depending on the work situation, needs of the candidate, accessibility of the item, and local industry and regional contexts) may also be included.

Codes of practice

Where reference is made to industry codes of practice, and/or Australian/international standards, it is expected the latest version will be used

Standards, codes, procedures and/or enterprise requirements

Standards, codes, procedures and/or enterprise requirements may include:

Australian and international standards, such as:

AS 2162.1-1996 Verification and use of volumetric apparatus - General - Volumetric glassware

AS 2252 Biological safety cabinets

AS 3753-2001 Recommended practice for chemical analysis by ultraviolet/visible spectrophotometry

AS ISO 17025-2005 General requirements for the competence of testing and calibration laboratories

AS/NZS 1269 Set:2005 Occupational noise management set

AS/NZS 1337 Eye protection

AS/NZS 2161 Set:2008 Occupational protective gloves set

AS/NZS 2210:1994 Occupational protective footwear

AS/NZS 2243.3:2002 Safety in laboratories - Microbiological aspects and containment facilities

AS/NZS 4501 Set:2008 Occupational clothing set

HB 9-1994 Occupational personal protection

Australian code of good manufacturing practice for medicinal products (GMP)

calibration and maintenance schedules

cleaning, hygiene, personal hygiene requirements

enterprise procedures, standard operating procedures (SOPs) and operating manuals

equipment startup, operation and shutdown procedures

European Union (EU)

Guide to physical containment levels and facility types

guidelines for small scale genetic manipulation work from the gene technology regulations

incident and accident/injury reports

instructions to comply with new legislation, standards, guidelines and codes

material, production and product specifications

National Association of Testing Authorities (NATA) Accreditation programs requirements

National Health and Medical Research Council (NHMRC)

National Registration Authority (NRA)

principles of good laboratory practice (GLP)

production and laboratory schedules

quality manuals and equipment and procedure manuals

quality system and continued improvement processes

safety requirements for equipment, materials or products and material safety data sheets (MSDS)

sampling procedures (labelling, preparation, storage, transport and disposal)

schematics, work flows and laboratory layouts

test procedures (validated and authorised)

Therapeutic Goods Regulations 1009

United States Food and Drug Administration (USFDA)

validated and authorised test methods

waste minimisation, containment, processing and disposal procedures

World Health Organisation (WHO)

Hazards

Hazards may include:

electric shock (e.g. electrophoresis power packs)

microbiological organisms and agents associated with soil, air, water, blood and blood products, and human or animal tissue and fluids

chemicals, such as acrylamide, temed, phenol and ammonium persulphate

mutagens, such as ethidium bromide, tumour promoters and cytotoxic materials

genetically altered organisms, transformed cultures and organisms

allergenic proteins

radioisotopes

transilluminators and other ultraviolet (UV) light sources

aerosols from broken centrifuge tubes and pipetting

sharps and broken glassware

flammable liquids and gases

cryogenics, such as dry ice and liquid nitrogen

disturbance or interruption of services

Safe work practices and hazard control measures

Safe work practices and hazard control measures may include:

ensuring access to service shut-off points

recognising and observing hazard warnings and safety signs

labelling of samples, reagents, aliquoted samples and hazardous materials

handling and storage of hazardous materials and equipment in accordance with labelling, MSDS and manufacturer's instructions

identifying and reporting operating problems or equipment malfunctions

cleaning and decontaminating equipment and work areas regularly using enterprise procedures

using personal protective clothing and equipment, such as gloves, safety glasses, coveralls, gowns, body suits and respirators

using containment facilities (PCII, PCIII and PCIV physical containment laboratories), containment equipment (biohazard containers, laminar flow cabinets, Class I, II and III biohazard cabinets) and containment procedures

reporting abnormal emissions, discharges and airborne contaminants, such as noise, light, solids, liquids, water/waste water, gases, smoke, vapour, fumes, odour and particulates to appropriate personnel

Equipment and instrumentation

Equipment and instrumentation may include:

pipettes, tubes and racks

heating blocks and polymerase chain reaction (PCR) thermal cyclers

swabs

centrifuges and shakers

electrophoresis tanks and power supplies

incubation cabinets for micro-organisms and cell culture

liquid nitrogen containers

autoclaves

water baths

waste containers

fumehoods

analytical instruments, such as spectrophotometers

Reagents

Reagents may include:

DNA, RNA and proteins

enzymes (restriction, ligation and polymerisation)

buffers

agarose, starch and polyacrylamide for electrophoresis gels

commercial kits for extraction and manipulation of DNA/RNA

phenol and chloroform

ethidium bromide

cell and culture media

DNA, and protein stains

specialised probe materials, such as radioactive, chemical and chemiluminescent labels

blotting membranes

chromatographic media

Molecular biology tests and procedures

Molecular biology tests and procedures may include:

generic skills:

sample digestion, extraction, filtration, separation, dialysis, precipitation and centrifugation

accurate and reliable use of micropipettes

application of aseptic techniques

labelling (e.g. digoxin, fluorescence, enzymes, radioactivity and antibodies)

production, labelling and use of DNA probes

preparation of competent bacterial cells

preservation and storage of samples (e.g. freezing)

extraction of nucleic acids:

isolation of genomic and plasmid DNA and RNA from samples, such as plants, bacterial suspensions, white blood cells, cheek cells, animal and plant tissue, cultured cells and forensic specimens

mini-prep and rapid method isolation of plasmid DNA

purification of nucleic acids and proteins:

purification of DNA using cesium gradients, commercial purification buffer kits and columns

purification of recombinant protein by chromatography

production of nucleic acids:

amplification of DNA by polymerase chain reaction

transformation with recombinant DNA

identification of transformed organisms with appropriate selection and analytical techniques, such as selective media and insertional inactivation

use of enzymes:

storage and handling of enzymes taking into account segregation, temperature, buffers and labelling to avoid wastage, denaturation and contamination

ligation

assistance with analysis of nucleic acids and proteins:

sequencing DNA

assaying of DNA purity and concentration using spectrometric analysis

electrophoresis of restriction enzyme digests of plasmid and genomic DNA using agarose gel

DNA sequencing by Sanger method

testing using restriction fragment length polymorphism (RFLP), probes and microsatellites

detection of protein products by measuring activity, including a range of immunological assays

assistance with hybridisations:

hybridisation to screen cDNA libraries

blotting (southern blots for DNA and Western blots for protein)

cloning:

cloning and sub-cloning of genes and fragments of DNA

applications of techniques:

PCR

methods to detect gene expression, such as RNA hybridisation, immunological techniques and radioactive labelling

testing DNA for sequence variation that is either causative of, or associated with, human disease

testing blood for the presence of viruses using the polymerase chain reaction

identification of species, such as bacterial contaminants

generating data for taxonomic and ecological investigations

Corrective action

Corrective action may involve:

purification

dilution

additional extraction steps

Records

Records may include:

test and calibration results

equipment use, maintenance and servicing history

photo images of gels, radioisotopes and digital images

chain of custody from sample to result

supplier certificates of analysis

quality control/analysis data

Occupational health and safety (OHS) and environmental management requirements

OHS and environmental management requirements:

all operations must comply with enterprise OHS and environmental management requirements, which may be imposed through state/territory or federal legislation - these requirements must not be compromised at any time

all operations assume the potentially hazardous nature of samples and require standard precautions to be applied

where relevant, users should access and apply current industry understanding of infection control issued by the National Health and Medical Research Council (NHMRC) and State and Territory Departments of Health