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.
The following two tabs change content below.
Kelly Grooms
Scientific Communications Specialist at Promega Corporation
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.
Latest posts by Kelly Grooms (see all)
- From Outside the Window to Inside the Cell: How Photography has Broadened Our View of the World - August 19, 2022
- Monkeypox—The Latest Zoonotic Virus Making Headlines - July 21, 2022
- PD-1 Blockade Treatment shows 100% Tumor Resolution in Mismatch Repair Deficient Rectal Cancer Patients - June 21, 2022
