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Contamination Control During PCR Reagent Preparation in the LPA Lab
Learning ObjectivesContamination Control During PCR Reagent Preparation in the LPA Lab
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During PCR Reagent preparation, if DNA fragments from the lab environment, such as a DNA template amplified in a previous qPCR experiment, enter the qPCR reaction or reagents (even in small quantities), they can be amplified during the reaction. This contamination and non-specific amplification can cause misleading results, such as false positives. Hence, contamination needs to be rigorously controlled.
Do’s and Don’ts for Contamination Control during PCR Reagent Preparation
Do’s
- Change shoes or use shoe covers, before entering the Reagent Preparation/ Master Mix Room.
- Always clean the work area properly before and after completing the work, using disinfectants.
- Keep master mix reagents in -20°C freezer.
- Label tools and instruments to be used only in Master Mix Room.
- Use 0.5 ml individual PCR tubes.
- Use a PCR hood.
- Decontaminate and calibrate pipettes regularly.
- Always prepare fresh disinfectant (1% sodium hypochlorite solution).
- Establish regular (e.g., weekly) and thorough laboratory cleaning protocols for floors, doors, walls.
Don’ts
- Tools and instruments used in Master Mix Room should not be used elsewhere.
- After working in other areas, do not come back and clean the reagent preparation area.
Resources
- GenoType MTBDR plus ver 2.0 Kit, Instructions for Use.
- GenoType MTBDRsl VER 2.0 Kit, Instructions for Use.
- GLI Training Package on LPA.
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