Ten Most Common Real-Time qRT-PCR Pitfalls

List of 10 possible reasons for an unsuccessful real-time PCR (Ambion inc.)

More detailed description can be found from this page.

Poor primer and probe design.
Using poor quality RNA.
Not using "master mixes".
Introducing cross-contamination.
Not using a "– RT" control.
Using an inappropriate normalization control.
Dissociation (melting) curves are not performed when using SYBR® Green.
Not setting the baseline and threshold properly.
The efficiency of the reaction is poor.
Using an inappropriate range for standard curves.

Last update 29-Jun-2006, Rating n/a of 0 votes.