Contamination Control During PCR Reagent Preparation in the LPA Lab

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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