10. Modify reaction buffer composition to adjust pH and salt concentration.

9. Titrate the amount of DNA polymerase.

8. Add PCR enhancers such as BSA, betaine, DMSO, nonionic detergents, formamide or (NH₄)₂SO₄.

7. Switch to hot-start PCR.

6. Optimize cycle number and cycling parameters, including denaturation and extension times.

5. Choose PCR primer sequences wisely.

4. Determine optimal DNA template quantity.

3. Clean up your DNA template to remove PCR inhibitors.

2. Determine the optimal annealing temperature of your PCR primer pair.
[Drum roll please]…and the number one thing you can do to improve your PCR results is:

1. Titrate the magnesium concentration.

Kelly Grooms

Kelly earned her B.S. in Genetics from Iowa State University in Ames, IA. Prior to coming to Promega, she worked for biotech companies in San Diego and Madison. Kelly lives just outside Madison with her husband, son and daughter. Kelly collects hobbies including jewelry artistry, reading, writing and knitting. A black belt, she enjoys practicing karate with her daughter as well as hiking, biking and camping.

• Kelly Grooms

  https://www.promegaconnections.com/author/kellygrooms/

  

  September 8, 2021

  New Cleared IVD Assay for Microsatellite Instability in Colorectal Cancer Aims to Help Identify Those with Lynch Syndrome
• Kelly Grooms

  https://www.promegaconnections.com/author/kellygrooms/

  

  June 22, 2021

  Questions Arise about TMB as a Predictive Biomarker for Immune Checkpoint Inhibitor Therapy
• Kelly Grooms

  https://www.promegaconnections.com/author/kellygrooms/

  

  June 9, 2021

  Improving SARS-CoV-2 Antibody Detection with Bioluminescence
• Kelly Grooms

  https://www.promegaconnections.com/author/kellygrooms/

  

  April 28, 2021

  There’s a Vaccine for That—Could mRNA Vaccines be Used to Prevent Cancer Recurrence?