Guidelines PCR analysis

Guidelines

• For starters, take a hands-on course on PCR and pipetting.
• Think about your experiment, plan and consult to avoid unnecessary or unusable runs.
• Buy what you need and do it together.
• Use best practice guidelines and relevant controls.
• Avoid optimization runs with relentless in-silico characterization of primers, probes and normalization genes.
• Do not store your work stocks of primers and probes separately, but together in 1 ep.
• Work in small volumes (10µl for normal reaction, 20µl for cloning), consider 384 well plates.
• Avoid duplicates as much as possible.
• Automation can reduce consumption of consumables.
• Set room temperature for your last cycle.

Course

Organise a hands-on course. Information is following.

Interesting links

• A beginner’s guide to RT-PCR, qPCR and RT-qPCR
• Good practice guide for the application of quantitative PCR (qPCR), LGC
• The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments.