G E Truett1, J A Walker, D G Baker. 1. Developmental Genetics Laboratory, Pennington Biomedical Research Center, Baton Rouge, LA 70808, USA.
Abstract
BACKGROUND AND PURPOSE: Efficient methods for detection and elimination of Helicobacter spp. infections are needed to facilitate the development of Helicobacter-free mouse colonies. We developed an inexpensive, high-throughput method for preparation of fecal DNA for Helicobacter polymerase chain reaction (PCR) assays. METHODS: Fecal DNA was prepared by heating fecal pellets to 95 degrees C for 10 minutes in an alkaline solution, then adjusting the pH by addition of Tris buffer. This solution is used for PCR assays without purification of DNA. We then tested fostering as a method of generating Helicobacter-free mice. Litters born to Helicobacter-positive dams were transferred to Helicobacter-negative foster dams on the first day of life. RESULTS: Fostered pups tested Helicobacter negative up to 89 days of age, whereas pups raised by Helicobacter-positive dams were all test positive by 19 days of age. CONCLUSION: These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies.
BACKGROUND AND PURPOSE: Efficient methods for detection and elimination of Helicobacter spp. infections are needed to facilitate the development of Helicobacter-free mouse colonies. We developed an inexpensive, high-throughput method for preparation of fecal DNA for Helicobacter polymerase chain reaction (PCR) assays. METHODS: Fecal DNA was prepared by heating fecal pellets to 95 degrees C for 10 minutes in an alkaline solution, then adjusting the pH by addition of Tris buffer. This solution is used for PCR assays without purification of DNA. We then tested fostering as a method of generating Helicobacter-free mice. Litters born to Helicobacter-positive dams were transferred to Helicobacter-negative foster dams on the first day of life. RESULTS: Fostered pups tested Helicobacter negative up to 89 days of age, whereas pups raised by Helicobacter-positive dams were all test positive by 19 days of age. CONCLUSION: These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies.
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