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Biochemistry

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Troubleshooting a PCR

BIOCHM-G@XELJ

You have designed a PCR to genotype a new human transgene-expressing mouse model. Your primers should exclusively amplify the transgene and your expected amplicon size is approximately 450 bp.

Unfortunately, when you evaluate your PCR using gel electophoresis, you discover that it has yielded several non-specific bands of both larger and smaller sizes in addition to your desired product.

Which of the following is the most plausible cause for the generation of these non-specific products?

A

There were not enough cycles.

B

The annealing temperature was too high.

C

There was not enough ${ Mg }^{ 2+ }$ added to the PCR reaction.

D

The annealing temperature was too low.