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.
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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, photography and knitting. She would like to be an avid runner, as evidenced by her growing collection of running gear and her single half-marathon finishers t-shirt.
Latest posts by Kelly Grooms (see all)
- Optimizing PCR: One Scientist’s Not So Fond Memories - July 31, 2019
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- For the Birds: Knitting Nests for Baby Birds Might Just Help Your Health To - March 29, 2019