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PCR

PCR Problems and Solutions

Problem:

PCR Contamination

PCR contamination is caused by DNA samples which

Solutions to PCR contamination:

Use Zero Aerosol Pipette Tips.

Prepare fresh new autoclaved water.

 

Primer Dimers

Primer dimers are caused by lack of binding of the primers to the template due to inefficient primer annealing to the template due to

Higher affinity binding of the primers to each other or themselves also causes primer dimers.

Primer dimers are seen as major band which migrates at less than 100bp on an agarose gel.

Solutions to Primer Dimer Formation:

Optimize annealing temperature

Design new primers which have lower self-annealing or self-hybridization potential.

 

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