Application Hints and Troubleshooting
This part of our website is intended to help those who are new to PCR to get started. It will also help anyone who wants to learn ways to enhance the basic PCR technique, for instance, to increase PCR yield and fidelity or to troubleshoot problems. Because of its length, the content has been broken up into several sections.
General Considerations
- Setting up the laboratory to avoid contamination
- Special equipment
- PCR reaction components
- Thermal cycling profile for standard PCR
- Properties of PCR enzymes
Factors to Consider in RT-PCR
- Choice of RT-PCR enzymes
- Choice of reverse transcription primers
- Preparation of template RNA
- Primer design
- RT-PCR procedures
Prevention of Carry-over Contamination
- Preventing carry-over contamination with uracil-DNA glycosylase
- Preventing carry-over contamination in RT-PCR
Troubleshooting