Guidelines PCR analyse

10 steps

• For starters, take practical training in PCR and pipetting. 

• Think about your experiment, plan and deliberate to avoid unnecessary or unusable runs. 

• Buy what you need and do it together. 

• Use best practice guidelines and relevant controls. 

• Avoid optimisation runs with thorough in-silico characterisation of primers, probes and normalisation genes. 

• Do not store your working 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 final cycle.

Training

• Provide practical training!

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.