AUG 14, 2016 9:01 PM PDT

PCR Primer Design Pointers

WRITTEN BY: Carmen Leitch

In this video from Thermo Fisher Scientific, a few tips for better PCR primer design are presented. PCR, Polymerase Chain Reaction, is a common and vital part of genetic studies, biomedical research and medicine in general.

Considerations important to the creation of good primers, such as primer length and melting temperature of the primers are highlighted in the video.

In PCR, the reaction mixture cycles through a series of temperatures. One of those, the annealing temperature, is also related to specificity and can be varied if needed for troubleshooting. For example, a PCR reaction that produces non-specific amplification products can likely tolerate a higher, more stringent annealing temperature. If no product is observed following the PCR reaction, the annealing temperature could be lowered as one way of trying to generate a successful reaction product.

Some pitfalls of PCR primers are shown too, such as hairpins that can form in primers that have sequences amenable to bending such that ends of the primer anneal to each other. Another thing to avoid are long sequences of the same base, or primers that have ends with a lot of cytosine (C) or guanine (G) bases.
About the Author
  • Experienced research scientist and technical expert with authorships on over 30 peer-reviewed publications, traveler to over 70 countries, published photographer and internationally-exhibited painter, volunteer trained in disaster-response, CPR and DV counseling.
You May Also Like
NOV 04, 2020
Plants & Animals
Scientists Rediscover "Lost" Chameleon Species in Madagascar
NOV 04, 2020
Scientists Rediscover "Lost" Chameleon Species in Madagascar
Voeltzkow’s chameleon was recently rediscovered after disappearing for more than 100 years. According to an articl ...
NOV 12, 2020
Chemistry & Physics
Can a Chemical Be Both Nurturing and Destructive?
NOV 12, 2020
Can a Chemical Be Both Nurturing and Destructive?
On August 4, a megascale explosion almost leveled half of Beirut, the capital city of Lebanon. At the center of the negl ...
DEC 11, 2020
Chemistry & Physics
Cosmic Expansion: Universe Stretches, We don't
DEC 11, 2020
Cosmic Expansion: Universe Stretches, We don't
Physicists believe that we exist in an expanding universe. But what does that mean? Do our habitats, bodies, and atoms & ...
MAR 26, 2021
Earth & The Environment
Iconic Bald Eagles are Thriving
MAR 26, 2021
Iconic Bald Eagles are Thriving
Once dangerously on the brink of extinction, bald eagle populations are thriving throughout the lower 48 states of the U ...
MAY 30, 2021
Space & Astronomy
New Discovery Casts Origin of Milky Way into Doubt
MAY 30, 2021
New Discovery Casts Origin of Milky Way into Doubt
A detailed cross-section of another galaxy, known as YGC 10738, has cast previous ideas of how the Milky Way was formed ...
JUL 16, 2021
Earth & The Environment
Melting Mountain Glaciers in Asia Release Greenhouse Gas
JUL 16, 2021
Melting Mountain Glaciers in Asia Release Greenhouse Gas
High mountain glaciers that are melting in the Tibetan plateau in Asia contain greenhouse gases and should be counted in ...
Loading Comments...