• Pre-PCR Reagent Preparation Supplies

• Pre-PCR Specimen Preparation Supplies

• Post-PCR Amplification and Detection Supplies

• PCR Laboratory Workflow and Good Laboratory Practice

The sample Laboratory Design below outlines the equipment needed for the AmplicorÒ PCR Test process.

• Pre-PCR

1. Dead-air Box.
2. Reagent Refrigerator.
3. Biological Safety Hood.
4. Standard Clinical Centrifuge and/or Microfuge.
5. Dry-heat Temperature Block.
6. Specimen Refrigerator.

• Post-PCR

1. Incubator.
2. Thermal Cycler.
3. Plate Washer.
4. Plate Reader and Printer.

These supplies and equipment must be dedicated to Reagent Preparation. Some items are not necessary for all AMPLICOR^® tests.

* Commercial liquid household bleach typically contains a concentration of 5.25% sodium hypochlorite. A 1:10 dilution of bleach will produce 0.5% sodium hypochlorite solution.

GeneAmpR is a registered trademark of Roche Molecular Systems, Inc.,licenced to the Perkin-Elmer Corporation.

MicroAmpR is a registered trademark of the Perkin-Elmer Corporation.

These supplies and equipment must be dedicated to Specimen Preparation.   Some items are not necessary for all AMPLICOR® tests.

*Commercial liquid household bleach typically contains a concentration of 5.25% sodium hypochlorite.  A 1:10 dilution of bleach will produce 0.5% sodium hypochlorite solution.

MicroAmpR is a registered trademark of the Perkin-Elmer Corporation.

Amplification and detection PE Applied Biosystems GeneAmp^® PCR System 9600 or System 2400 Thermal Cycler. Incubator (37°C ± 2°C). Microwell plate washer. Microwell plate reader.

Printer and paper.

GeneAmpR is a registered trademark of Roche Molecular Systems, Inc.,licenced to the Perkin-Elmer Corporation.

In addition to the standard biosafety laboratory practices and good laboratory protocol, the following considerations should apply specifically to AMPLICOR® PCR Tests and should be monitored in order to ensure the integrity of the AMPLICOR® Test Kits.   For equipment requirements, procedural information and storage and handling requirements for all AMPLICOR® components, please refer to the individual package insert for each assay.

• Always work in a one-way direction from Pre-PCR to Post-PCR to avoid carryover contamination from amplified products.

• Post-PCR should be kept as far away as possible from Pre-PCR to avoid aerosol contamination.

• The working surface in each area must be decontaminated with 0.5% sodium hypochlorite followed by 70% ethanol before performing assay procedures in that area.

• Specimens must be stored separately from reagents so as not to contaminate open reagents.

• When handling material containing DNA/RNA or Amplicon, always use a pipettor with a plugged (aerosol-barrier) tip or positive displacement tip.   Post-PCR pipettors must never be used in Pre-PCR.

• To avoid possible aerosol contamination, all centrifuges should be kept at a distance from areas where the operator is preparing Master Mix and controls and adding prepared specimens to Master Mix.

• To avoid specimen contamination from seepage into poorly capped tubes, water baths are to be avoided; dry baths or dry heat blocks, if needed, are preferable.

• Incubator temperature should be kept stable.  Traffic in and out of an incubator should be reduced to a minimum.

• Lab coats must be worn in all areas. The coat worn in Post-PCR must never be worn in Pre-PCR!

• Gloves must be worn at all times for operator safety as well as for control of contamination from one area to another:
  • Gloves must be changed before moving to the next work area
  • Gloves worn in the specimen preparation area must never be worn in the reagent preparation area
  • Gloves worn in Post-PCR must never be worn in Pre-PCR