Primer-dimers; note everything is less than 100 bp in size
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| Seen on an UV transilluminator (blue band is bromophenol) |
Wednesday, November 18, 2020
Polymerase chain reaction and agarose gel electrophoresis of DNA
Electrophoresis run (blank) without PCR product, only control ('ladder') DNA.
Monitoring the progress of electrophoresis with bromophenol blue, which migrates at the same rate as a 150-300 bp DNA fragment
Often, one is not sure of the optimal temperature of a primer to anneal; in which case, trial and error at several temperatures is done. Modern thermocyclers allow different temperature for each row of wells (gradient PCR). The temperature with most product formation (thickest band) is selected.
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