Preventing Contamination; Compatible Consumables - Applied Biosystems 7900HT User Manual

Chapter 4 Operating the Instrument

Preventing Contamination

Contamination
and the
5´ Nuclease
Assay
General PCR
Practices

Compatible Consumables

Consumables for
Use with
the 7900HT
Instrument
4-6 Applied Biosystems 7900HT Fast Real-Time PCR System and SDS Enterprise Database User Guide
September 1, 2004 11:39 am, CH_Basic.fm
PCR using the 5´ nuclease assay requires special laboratory practices to avoid false
positive amplifications (Kwok and Higuchi, 1989). The assay's logarithmic
amplifications can potentially amplify a single DNA molecule (Saiki et al., 1985;
Mullis and Faloona, 1987).
Observe the following guidelines when assembling optical plates or loading
TaqMan ® Low Density Arrays for use on the 7900HT instrument:
• Wear a clean lab coat (not previously worn while handling amplified PCR products
or used during sample preparation) and clean gloves when preparing samples for
PCR amplification.
• Change gloves whenever you suspect they are contaminated.
• Maintain separate areas, dedicated equipment, and supplies for:
– Sample preparation
– PCR setup
– PCR amplification
– Analysis of PCR products
• Never bring amplified PCR products into the PCR setup area.
• Open and close all sample tubes carefully. Try not to splash or spray PCR samples.
• Keep reactions and components capped as much as possible.
• Use positive-displacement pipets or aerosol-resistant pipet tips.
• Clean lab benches and equipment with 10% bleach solution.
Table 4-1 Compatible Consumables
Consumable Type
ABI P ™
RISM
384-Well Optical
Reaction Plates
DRAFT
Compatible Seals
Use these...
• ABI P ™ Optical Adhesive Covers
RISM
(quantity 100)
• Optical Adhesive Covers (quantity 25)
Not these...
384-Well Septa