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PCR Lab Design Considerations

The document outlines guidelines for setting up a molecular laboratory with three separate work areas to prevent contamination: 1) reagent preparation, 2) specimen preparation, and 3) amplification/detection. Key recommendations include using unidirectional workflow from clean to dirty areas, separating areas physically and with air pressure controls, and following strict protocols for equipment, supplies, and personal protective equipment to minimize risk of contamination. Regular decontamination of surfaces, equipment, and the laboratory environment is also emphasized.

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Jihad Elias Chahla
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177 views

PCR Lab Design Considerations

The document outlines guidelines for setting up a molecular laboratory with three separate work areas to prevent contamination: 1) reagent preparation, 2) specimen preparation, and 3) amplification/detection. Key recommendations include using unidirectional workflow from clean to dirty areas, separating areas physically and with air pressure controls, and following strict protocols for equipment, supplies, and personal protective equipment to minimize risk of contamination. Regular decontamination of surfaces, equipment, and the laboratory environment is also emphasized.

Uploaded by

Jihad Elias Chahla
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPT, PDF, TXT or read online on Scribd
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Molecular Laboratory

Design
Setting Up a Molecular
Laboratory
• Mechanical barriers to prevent contamination
• Spatial separation of pre- and post-amplification work areas
• Area 1 – Reagent preparation
• Area 2 – Specimen preparation
• Area 3 – Amplification/product detection
• Physically separated and, preferably, at a substantial distance
from each other
Unidirectional Flow
• Both personnel and specimens
• Amplification product-free to product-rich
• Remove PPE before leaving one area
• Avoid or limit reverse direction
• Reusable supplies in the reverse direction need to be bleached.
Features of the 3 Areas

• Each area has separate sets of equipment and supplies


• Refrigerator/freezer (manual defrost)
• Pipettes, tips, tubes, and racks
• Centrifuge, timers, vortex
• Lab coat (color-coded), disposable gloves, safety glasses, and other
PPE
• Cleaning supplies
• Office supplies
• Ventilation system
• Dead air box with UV light – serves as a clean bench area
Features of the 3 Areas

• Air pressure
• Reagent Prep and Specimen Prep – Positive
• Postamplification - Negative
• Reagent Prep – Single entrance, reagents used for amplification
should not be exposed to other areas
• Specimen Prep – Specimens should not be exposed to post-
amplification work areas
Important Details

• Use of positive displacement pipettes and disposable filtertip


pipette tips
• Avoid production of aerosols when pipetting
• Use of sterilized single-use plasticware
• Minimizes the risk of amplicon carry-over on clothing, hair
and skin
• Hairnet
• Dedicated safety glasses
• Disposable labcoat/gown
• Gloves
• Shoe covers
More Important Details

• Use of nuclease free or autoclaved water


• Aliquot oligonucleotides – multiple freeze thaws will cause
degradation
• Always include a blank control to check for contamination
• Wipe test
• Monthly
• Detect and localize the contamination
• Identify the source of the contamination
Decontamination Approaches

• Clean the work area & equipments routinely


• Clean the PCR workstation at the start and end of each work day/run (UV
light, 70% ethanol, fresh 10% sodium hypochlorite, DNA Away)
• Clean the exterior and interior parts of the pipette
• Clean the equipment
• Clean the doorknobs, handle of freezers
Other Considerations

• Temperature and humidity requirements


• Exhaust ventilation
• Water quality
• Back-up power system

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