Question

A researcher is doing a PCR-based experiment using the following temperature profile: • Melt for 1 minute at 94°C • Anneal for 30 seconds at 37°C Extend for 30 seconds at 70°C Unfortunately, her gels show way too many bands. Her primers have perfect complementarity to the target sequence. Give a simple way to reduce the number of bands.

Answer 1

She can increase the extension time and annealing time .

Annealing time should be 1 Min and extension time can be 2 min minimum .

Small annealing time can lead to improper annealing and thus many length chain can be synthesized.

If extension time is also less then it can be also produced different bands because different extension can be happened.